Enhancement of motility by treating spermatozoa with an antioxidant solution (Sperm-Fit) following ejaculation

Oxygen radical generation is known to be detrimental to sperm function, especially motility, through the lipid peroxidation of the membranes. Generation of reactive oxygen species can be induced by leukocyte contamination, sperm centrifugation and the presence of abnormal spermatozoa with excess residual cytoplasm. This study aims to evaluate the effect on sperm motility of incubation in an antioxidant-containing solution, during liquefaction and centrifugation. Thirty semen samples were each divided into two equal parts: one mixed with Tyrode's solution, the other with a salt solution containing antioxidants (Sperm-Fit; Ellios Bio-Media, Paris, France). All the procedures were identical in the two groups. The ratio of leukocytes to spermatozoa was significantly correlated with the motility after liquefaction and after a 24 h incubation in routine in-vitro fertilization (IVF) medium and with the number of motile spermatozoa recovered after Percoll preparation. Moreover, when this ratio was > or = 0.2, all motility parameters were lowered. Incubation with Sperm-Fit allowed a higher percentage of motility after Percoll preparation when the ratio was > or = 0.2 (48 +/- 5% versus 41 +/- 6% for Sperm-Fit and Tyrode's solution respectively; P < 0.05) and a greater number of motile spermatozoa recovered after Percoll preparation, whatever the ratio (3.2 +/- 1.0 x 10(6) versus 2.4 +/- 0.7 x 10(6) for Sperm-Fit and Tyrode's solution respectively when ratio > or = 0.2; 18.1 +/- 3.4 x 10(6) versus 14.4 +/- 2.9 x 10(6) for Sperm-Fit and Tyrode's solution respectively when ratio < 0.2; P < 0.05). These results show that incubation with antioxidants during liquefaction and centrifugation increases recovery of motile spermatozoa.     

Microsatellite deletion mapping on chromosome 10q and mutation analysis of MMAC1, FAS, and MXI1 in human glioblastoma multiforme

The aim of this study was to determine the relationship between calcium ionophore A23187-induced acrosome reaction (AR) and sperm fertilizing ability. Semen samples remaining after preparation for standard IVF were studied in 109 patients who had sperm concentrations > or =20 x 10(6)/ml. Ionophore-induced AR was performed on motile spermatozoa selected by centrifugation on a Percoll gradient. Semen analysis was performed using standard methods. Patients with higher (>50%, n = 76) fertilization rates had significantly higher ionophore-induced AR than patients with lower (<50%, n = 33) fertilization rates (49 +/- 14 versus 38 +/- 21%, P < 0.05). When the data from all patients were analysed by logistic regression, only the percentage sperm motility in insemination medium and ionophore-induced AR were significantly related to fertilization rates. Similar results were also obtained when the data from a subgroup of patients with poor (<15% normal) sperm morphology were analysed. However, when patients with normal sperm morphology > or =15% were analysed separately, only sperm count and the percentage of spermatozoa with progressive motility in semen were significantly related to fertilization rates. In conclusion, ionophore-induced AR was significantly related to fertilization rates in vitro mainly in patients with teratozoospermic semen. Tests for ionophore-induced AR may provide additional information about sperm fertilizing ability but may not indicate specific defects of the physiological AR.     

Comparison of various methods of processing human cryopreserved-thawed semen samples

We compared the efficacy of various methods of processing cryopreserved-thawed samples for the recovery of functionally adequate spermatozoa as assessed by the response to the sperm stress test (SST), an index of temperature activated sperm membrane lipid peroxidation, and immediate and delayed changes in sperm viability and motion parameters. Donor semen samples (n = 28) were cryopreserved-thawed and divided into six equal parts, one part was used as control and the remaining parts were used to compare five methods of sperm processing as follows: direct Percoll gradient processing, washing by one-step or stepwise addition of the washing medium followed by Percoll processing, and washing by one-step or stepwise addition of the washing medium. Additional samples (n = 10) were evaluated for the immediate and delayed (6 h at 37 degrees C) impact of one-step and stepwise washing (without Percoll separation). Compared with wash-only methods, samples processed using Percoll had a significantly higher SST score (P = 0.001), motility, rapid spermatozoa (>50 microm/s), curvilinear velocity and motility index (P < 0.001). Comparing various Percoll methods, direct Percoll processing resulted in the highest number of motile spermatozoa recovered (P < 0.00001) and a higher SST score based on curvilinear velocity (P = 0.001). Stepwise washing gave a significantly higher number of motile spermatozoa (P < 0.001) but with a significantly lower SST score based on the concentration of motile spermatozoa (P = 0.001), motility (P = 0.001) and motility index (P = 0.01). Sperm viability and motion parameters after 6 h of incubation showed no difference between one-step and stepwise washing. In conclusion, compared with wash-only methods, Percoll processed samples resulted in the recovery of spermatozoa with superior quality as assessed by SST and motion analysis. One-step washing of the samples gave an overall comparable recovery compared to the samples prepared stepwise. Having significantly higher SST scores, similar viability and the maintenance of motility, one-step washing may be a better method of processing thawed samples than the stepwise washing.     

Simultaneous assessment of sperm chromatin condensation and morphology before and after separation procedures: effect on the clinical outcome after in vitro fertilization

OBJECTIVE: To look for correlations between acridine orange (AO) staining and semen parameters before and after sperm separation procedures and to assess whether the AO test predicts fertilization or pregnancy outcomes after standard IVF and intracytoplasmic sperm injection. DESIGN: Prospective study that simultaneously assesses sperm morphology and nuclear protein maturity on a cell-by-cell basis before and after preparative procedures. SETTING: University teaching hospital. PATIENT(S): Men (n = 140) undergoing diagnostic semen analysis. MAIN OUTCOME MEASURE(S): Acridine orange fluorescence of sperm nuclei, semen parameters, IVF outcome. RESULT(S): In unprocessed samples, 90% of sperm with normal heads displayed green fluorescence (mature nuclear protein); significantly lower percentages of green fluorescence were observed in sperm with abnormal heads. The percentage of mature normal sperm in the specimen correlated with motility. Sperm maturity after swim-up or Percoll gradient was significantly improved for sperm with normal or abnormal heads. The percentage of mature normal sperm correlated with motility after either Percoll or swim-up. Neither the percentages of mature nuclei nor mature normal nuclei correlated with fertilization or pregnancy outcome. CONCLUSION(S): Nuclear protein maturation correlates with sperm motility and morphology. Because morphologically normal and motile sperm are more mature, separation procedures should generate a population of sperm with the highest fertilization capacity. Acridine orange staining, however, did not predict fertilization efficiency or pregnancy outcome in IVF cycles.     

Comparison of single- and two-layer Percoll separation for selection of motile spermatozoa

OBJECTIVE: Sperm recovery using a single-layer Percoll procedure is significantly better than using the swim-up technique for infertile men and patients with normal sperm characteristics; however, in normal men results have been contradictory. Some studies have shown further improvement in semen quality with multiple layers. Therefore, this study compared the effect of single-layer and two-layer Percoll procedures on sperm characteristics of normozoospermic men. METHODS: Semen specimens from 10 normal donors were processed by layering 1 mL of the liquefied ejaculate on a single layer of 80% Percoll or on a two-layer (47% and 90%) Percoll gradient. Computer-assisted semen analysis was done to examine total motile sperm, percentage of recovery of motile cells, percent motility, curvilinear velocity, linearity, and amplitude of lateral head displacement. Each specimen was evaluated by the hypo-osmotic swelling (HOS) test, bovine cervical mucus penetration test, viability (eosin-nigrosin stain), and sperm morphology (World Health Organization and Kruger's strict criteria). RESULTS: Specimens processed with the two-layer Percoll procedure had significantly better recovery of spermatozoa, and significantly better percentage motility, linearity, amplitude of lateral head displacement, percentage tail swelling, and percentage viability than those separated on single-layer Percoll. Results for sperm morphology using WHO and Kruger's criteria were similar between the two methods (P = 0.92 for both sets of criteria). CONCLUSIONS: In normozoospermic men, the two-layer Percoll separation procedure significantly improves semen characteristics compared with separation on a single layer.     

Spermatozoa selection by the swim-up procedure and two-layer percoll gradient centrifugation

The efficiency of the swim-up procedure and two-layer Percoll gradient centrifugation in procession of spermatozoa was assessed in ejaculates from 47 infertile men. A significantly higher total number of spermatozoa was harvested from Percoll gradients than from the swim-up procedure, the loss rates in concentration being -13.6 +/- 6.4% and -70.8 +/- 5.8%, respectively (p < 0.0001). Recovery in per cent motility was significantly higher after the Percoll gradient than after the swim-up procedure (34.8 +/- 10.2% versus -10.4 +/- 17.2%, p < 0.05). No significant difference was noted between the mean motility grades of the final solutions obtained by the two methods (2.7 +/- 0.2 and 2.0 +/- 0.4, respectively, p > 0.05). When evaluation was conducted within three initial fresh sample concentration categories such as severe oligospermia (lower than 5 x 10(6)/ml), moderate oligospermia (5 to 10 x 10(6)/ml) and mild oligospermia (higher than 10 x 10(6)/ml), the Percoll technique recovered significantly higher number of spermatozoa than the swim-up procedure through all concentration categories (p < 0.05 for each range). Despite being statistically insignificant, Percoll gradients produced final spermatozoa pools with higher per cent motility and motility quality within all concentration ranges. The results suggest that the Percoll gradient centrifugation should be the preferred selection method regardless of the initial fresh sample concentration.     

Assessment of sex chromosome ratio and aneuploidy rate in motile spermatozoa selected by three different methods

Using three-colour fluorescence in-situ hybridization, sex chromosome ratios and frequencies of diploidy and disomy for chromosomes X, Y and 18 were compared in spermatozoa of good and poor motility after separation by swim-up, glass-wool and two-layer discontinuous Percoll methods. Semen samples were collected from seven normal males aged 26-31 years. A minimum of 6000 sperm nuclei per sample were evaluated for each chromosome for a total of 308,432 sperm nuclei. Hybridization efficiency was 99.8%. A slight change in the ratio of X- to Y-bearing spermatozoa was noted after Percoll separation (from 49.3:49.5 to 50.0:48.9; P = 0.036 and P = 0.046), but not after separation by the other two methods. We did not observe significant differences in the disomy rates for sex chromosomes or chromosome 18 or in the diploidy rate between spermatozoa with good and poor motility after separation by any of the three methods. Our data indicate that separation of motile spermatozoa does not alter the ratio of X- to Y-bearing spermatozoa to a degree that represents sex chromosome selection.     

Study of aneuploidy in normal and abnormal germ cells from semen of fertile and infertile men

This study was undertaken with the aim of investigating the cytogenetic constitution of normal as well as abnormal spermatozoa and immature germ cells found in semen of normal men and infertile patients. A specific protocol of double in-situ hybridization for chromosomes 1 and 17 based on colorimetric detection of the hybridization signals (ISH) and brightfield microscopy analysis of cellular morphology was applied. Also the influence of paternal age on sperm aneuploidy was investigated. We found that, at least in the age range analysed (28-54 years) and for semen of good quality (total normal motile counts above 10 x 10(6)) (n = 17), paternal age has no influence on baseline rates of sperm aneuploidy. However, with decreasing semen quality (total normal motile sperm counts below 5 x 10(6)) (n = 6) significantly higher rates of sperm aneuploidy for autosomes 1 and 17 were scored (0.8 versus 1.43%) (P < 0.001). Regardless of the type of semen analysed, a number of morphologically abnormal spermatozoa were found to be hyperhaploid or diploid in a high percentage of cases (20 and 10% respectively). The same was found for immature germ cells (aneuploidy rate of 18%). We conclude that in infertile men with poor quality semen a direct relationship may exist between the impairment of the spermatogenesis process (as reflected by an increased production of morphologically and cytogenetically abnormal germ cells) and rates of baseline aneuploidy occurring in normal spermatozoa. Infertile couples undergoing assisted reproduction treatment need to be counselled about the risk of using spermatozoa which may carry higher rates of non-disjunction for different chromosomes. While sperm hyper- or hypohaploidy for some chromosomes (X,Y) implies counselling couples about the risk of abnormal phenotype in their offspring, most autosomal sperm aneuploidies probably translate only into lower rates of embryo fertilization and survival.     

Selective capacity of glass-wool filtration for the separation of human spermatozoa with condensed chromatin: a possible therapeutic modality for male-factor cases?

PURPOSE: The aim of this study was to evaluate chromatin condensation of human spermatozoa following swim-up compared to glass-wool separation. Semen aliquots from men attending an andrological outpatient clinic were processed by means of a swim-up procedure and glass-wool filtration. Chromatin condensation was recorded using aniline blue staining and results were reported according to color intensity of stained sperm heads. Morphometric measurements of sperm heads were performed on stained sperm samples. RESULTS: Glass-wool filtration resulted (i) in a significantly higher total motile sperm count (P < 0.0005) compared to swim-up and (ii) in a significantly higher percentage of normal chromatin-condensed spermatozoa compared to the ejaculate (P < 0.01). CONCLUSION: In contrast, comparing swim-up to the ejaculate, the percentage of matured nuclei (unstained spermatozoa) retrieved following swim-up was significantly lower (P < 0.005). Glass-wool filtration separates human spermatozoa according to motility and size of the sperm head. The size of the sperm head closely correlated with the chromatin condensation quality.     

Electrodermal level asymmetry in a unilateral brain-damaged patient

To compare the effectiveness of various sperm preparation methods, we examined sperm concentration, percentage of progressive motility, recovery rate, and various motion characteristics in 32 semen samples after Percoll and IxaPrep preparations. Except for sperm concentration, which was much higher after IxaPrep preparation, no statistical differences existed between these two methods in terms of motile sperm concentration (MSC), recovery rate, mean curvilinear velocity (VCL), mean straight-line velocity (VSL) and mean amplitude of lateral head displacement (ALH). Among 22 samples that were allocated in a 37 degrees C, 5% CO2 incubator, the mean MSC after 3 hours was significantly higher following IxaPrep preparation than following Percoll preparation (63.2 x 10(6)/mL vs. 42.8 x 10(6)/mL, p < .03). VCL and VSL were much higher after IxaPrep preparation than after Percoll preparation, both at 3 hours (p < .01) and at 24 hours (p < .03). ALH was also higher after IxaPrep preparation than after Percoll preparation, and it was statistically significant after 3 hours of incubation (p < .03). We conclude that the IxaPrep gradient may be adopted as the preferred method to the Percoll gradient in preparing spermatozoa for assisted reproduction because of the spermatozoa's persistent and better motion activities after incubation.     

Comparison of sperm separation methods: effect on recovery, motility, motion parameters, and hyperactivation

OBJECTIVE: To compare the Enhance (Percoll; Conception Technologies, San Diego, CA) and PureSperm (Gen X International, Madison, CT) sperm preparation methods with respect to recovery (percentage of motile sperm), motility (%), path and progressive velocities (microm/s), and hyperactivation (%). DESIGN: Comparison of sperm processing methods. SETTING: University medical center-based clinical andrology laboratory and infertility program. PATIENT(S): Twenty-five men who presented for semen analysis. INTERVENTION(S): Each of 25 semen specimens were divided and each aliquot was prepared using two different density gradient centrifugation methods. MAIN OUTCOME MEASURE(S): The motile sperm recovery, percent motility, motion parameters, and percent hyperactivation were measured for each semen specimen (n=25) before and after separation with the use of the two methods. RESULT(S): There was no difference in the percent motility and motile count between specimens prepared with Enhance (Percoll) and PureSperm and fresh specimens. Statistically significant differences were found (fresh versus test) in the velocities and in hyperactivation (PureSperm only), and no differences were found between the processing methods. CONCLUSION(S): PureSperm appears to be as effective as Percoll (Enhance) for the recovery of good, progressively motile sperm for use in IUI or other assisted reproductive techniques.     

Optimal utilization of cryopreserved human semen for assisted reproduction: recovery and maintenance of sperm motility and viability

PURPOSE: Our purpose was to evaluate sperm motility and viability and the maintenance of these parameters in already cryopreserved semen samples following repeated freezing/thawing cycles. METHODS: Human spermatozoa were subjected to five cycles of cryopreservation/thawing. Recovery of sperm motility and viability and the proportion of viable nonmotile sperm were determined up to 6 hr after thaw. RESULTS: Sperm motilities (prefreeze motility, 70.1%; n = 9 samples) after each of five freeze/thaw cycles were 24.4, 8.0, 3.5, 1.5 and 1.8%. The recovery of sperm viability was higher than that of motility after each cycle: 39.1, 25.3, 22.6, 17.8, and 16.5%. Recoveries of motility and viability were improved if the thawed samples were left in the original cryopreservation medium prior to refreezing vs. if a washing/ resuspension step was included. The recovery of sperm motility in the first thawing cycle was indicative of the expected motile sperm recovery in the second thawing cycle. CONCLUSIONS: Cryopreserved semen that is intended to be reused in future assisted reproduction treatments should be thawed only once and aliquoted in the original freezing medium before refreezing. The recovery of sperm motility and viability in the second thawing cycle, thus the applicability of the sample in conventional in vitro fertilization or intracytoplasmic sperm injection may be anticipated in > 90% of the samples. In view of intracytoplasmic sperm injection it is important that sperm viability is maintained better than motility; after the first, second, and third thawing cycles the ratios of motile:nonmotile viable sperm were 1:1, 1:4, and 1:7, respectively.     

Variation in recovery of motile sperm after preparation by a simple Percoll gradient technique

PURPOSE: Numerous techniques have been used to prepare sperm for assisted reproduction technology. Density-gradient centrifugation with Percoll is becoming a method of choice. This study reviewed the results of a simple two-layer discontinuous Percoll gradient. METHODS: We reviewed retrospectively results obtained from 208 semen specimens prepared for in vitro fertilization by a discontinuous Percoll gradient. RESULTS: Overall results (mean +/- SD) were a recovery of 21 +/- 16% of total sperm, recovery of 38 +/- 30% of motile sperm, and a motility in the final sperm preparation of 88 +/- 10%. Specimens with higher initial concentrations had higher motility in the final preparation. Higher total recovery and higher motility in the final preparation were found for specimens with a higher initial motility. CONCLUSIONS: This simple two-layer Percoll technique is rapid and inexpensive and reliably produces a final sperm preparation with desirable characteristics. Even though specimens with poor initial parameters yielded final preparations with excellent characteristics, fertilization rates were still related to the initial semen parameters.     

Relationship of bacteriological characteristics to semen indices and its influence on fertilization and pregnancy rates after IVF

BACKGROUND: To determine if routine semen culture is useful in asymptomatic couples joining an in-vitro fertilization (IVF/ET) program. METHODS: Bacterial cultures and semen analysis according to WHO recommendations were performed on semen samples obtained before oocyte recovery from 88 asymptomatic couples undergoing IVF during a 7-month period. RESULTS: In 46 cultures at least one kind of microorganisms could be isolated. Forty-two cultures either contained bacterias regarded as normal skin flora (n = 14) or showed no growth of microorganisms (n = 28). No differences were found in sperm concentration, total sperm count and sperm morphology between the semen samples with positive bacteriology and those with negative culture results. Sperm motility was decreased before Percoll preparation if microorganisms were present. Positive culture results had no effect on either fertilization or pregnancy rates. CONCLUSIONS: These observations suggest that bacteriospermia is not associated with abnormal sperm function after Percoll preparation or adverse IVF outcome.     

Synthesis and antibacterial evaluation of novel water-soluble organic peroxides

Various compounds have been used in the attempt to improve sperm motility, including pentoxifylline (PF), a methylxanthine derivative. It has been postulated that PF, being a phosphodiesterase inhibitor, increases sperm kinematic parameters and the number of spermatozoa exhibiting hyperactivated motility by raising the intracellular content of cAMP, a molecule involved in the generation of sperm energy. However, it has not been clarified whether the biological effects of PF on sperm motility correlate with its ability to increase intracellular cAMP levels. To examine this relationship, the kinematic parameters, hyperactivation, and intracellular cAMP content were evaluated in motile spermatozoa, obtained by discontinuous Percoll gradient and swim-up from 21 normozoospermic semen samples, incubated without and with PF for 0, 1, 2, and 4 h. PF increased beat cross frequency after 1 and 2 h of incubation, curvilinear velocity and lateral head displacement (ALH) after 4 h, and hyperactivation after 1, 2, and 4 h, and decreased linearity (LIN) after 1 h of incubation. The intracellular cAMP content of spermatozoa incubated with PF increased at all time-points examined. Both intracellular cAMP content and increase in hyperactivation in response to PF decreased with the length of incubation. In the absence of PF, cAMP content was unchanged and was correlated significantly only with ALH and the percentage of spermatozoa with hyperactivated motility. Following incubation with PF, cAMP content correlated with hyperactivation and all sperm kinematic parameters, with the exception of LIN and straightness. These findings suggest that the beneficial effects of PF on sperm kinematic parameters and hyperactivation are related to its ability to increase intracellular cAMP content.     

Environmental, management, and genetic factors affecting semen production in Holstein bulls

The objective of this study was to evaluate the importance of environment, management, physiological status, and genetics on semen quality (volume of the ejaculate, sperm concentration, sperm motility, number of sperm, and number of motile spermatozoa per ejaculate) of Canadian Holstein bulls. For this purpose, semen production data from 198 bulls were analyzed using mixed linear models. Young bulls (up to 30 mo old) and mature bulls (between 4 and 6 yr old) were analyzed separately. Semen characteristics generally improved significantly with age of young bulls. Season significantly affected all semen traits in young bulls but did not significantly affect volume and sperm motility of mature bulls. Performance was better in winter than in summer. The highest numbers of motile spermatozoa per ejaculate were obtained with intervals of at least 4 to 5 d between collections. Although the bull handler and semen collector caused less than 10% of the variance, the collection team significantly affected semen volume, number of sperm, and number of motile sperm per ejaculate for both growing and mature bulls. Heritabilities for volume, concentration, sperm motility, number of sperm, and number of motile sperm per ejaculate were, respectively, 0.24, 0.52, 0.31, 0.38, and 0.49 for young bulls and 0.44, 0.36, 0.01, 0.54, and 0.64 for mature bulls. Repeatability of semen traits varied from 0.41 to 0.64. Genetics, management, and environmental factors clearly contribute to semen production in Holstein bulls.     

Improved motile sperm recovery by a hyperosmotic Percoll gradient

PURPOSE: Our purpose was to investigate whether a new, relatively hyperosmotic Percoll gradient, Enhance-S, can improve total motile sperm recovery rates compared with the commonly used Percoll gradient Perception. METHODS: Semen specimens from each of 17 donors were divided into two equal aliquots. One part was washed using Percoll Perception, while the other was prepared using Percoll Enhance-S. RESULTS: Compared to the unwashed specimen, sperm motion characteristics (motility and velocity) improved significantly after Percoll separation using either the Perception or the Enhance-S gradient. There was no difference in motility or velocity in spermatozoa recovered after wash with either of the two preparations. However, the total motile sperm recovery was significantly higher using the Percoll Enhance-S gradient than with the Percoll Perception gradient (P < 0.0024). CONCLUSION: The new Percoll Enhance-S gradient provides significantly more total motile sperm than the Percoll Perception gradient.     

Rotational digital subtraction carotid angiography: technique and comparison with static digital subtraction angiography

To compare the efficiency of sperm preparation between the two-layer Percoll gradient and mini-Percoll methods, 50 normal and 33 abnormal semen samples from male partners of infertile couples were studied. The number of recovered spermatozoa, percentage of motility, percentage of normal morphology, and their survival at 24 and 48 hours were assessed. Both Percoll gradient techniques resulted in a significantly higher percentage of motility and percentage of normal morphology compared with the original semen samples (p < 0.0001). The two-layer Percoll gradient showed a higher sperm recovery than the mini-Percoll method (p < 0.001), but the latter resulted in a higher percentage of motility (p > 0.001) and a higher sperm survival rate at 24 hours (p < 0.05) than the former, regarding normal semen samples. These differences did not appear with abnormal semen samples when analyzed as a group. Considering each of the abnormal parameters separately, sperm recovery was significantly higher after the two-layer Percoll gradient in the case of astheno- and teratozoospermia (p < 0.05), but sperm survival at 48 hours was higher after the mini-Percoll gradient in the case of teratozoospermia (p < 0.05). It is concluded that both the two-layer Percoll gradient and mini-Percoll method can be used effectively for sperm preparation. The former yields a higher sperm recovery, but the latter should be considered regarding teratozoospermic samples and semen samples of very low volume.     

Strategies in frozen donor semen procreation

Data were analysed from 710 couples who had been assessed to determine the effectiveness and the drawbacks of three different methods of insemination using frozen donor semen. Intracervical insemination (ICI) was the first method used when the women had no tubal disorder: 255 pregnancies were achieved in a total of 2558 cycles (10%). Intrauterine insemination (IUI) associated with ovarian stimulation resulted in 152 pregnancies over 966 cycles (16%). In-vitro fertilization (IVF) was proposed after approximately 12 insemination failures using either of the other methods or when the initial gynaecological examination had revealed abnormalities such as tubal occlusions; 48 pregnancies were obtained in 262 cycles (18.3%). The pregnancy rate using ICI was significantly higher when two inseminations were performed per cycle, compared with one insemination per cycle (12.3 versus 7%, P < 0.001). The number of motile spermatozoa per straw was correlated with the pregnancy rate when using ICI, rising from 9% with < 4 x 10(6) motile spermatozoa to 13.8% with 4-8 x 10(6) and 17.2% with > 8 x 10(6). No relationship was found between the number of motile spermatozoa and the pregnancy rate using IUI and IVF. The incidence of primary ovulatory disorder was higher among women whose husbands were oligozoospermic than among those whose husbands were azoospermic (19 versus 9%, P < 0.01), but ovarian stimulation improved the fecundity of subfertile women. The outcome of pregnancies was also analysed for the three methods. From these data, strategic plans have been proposed to maximize the pregnancy rate for women undergoing therapeutic donor insemination with frozen semen.     

Investigation of numbers and motility of spermatozoa in reproductively active and socially suppressed males of two eusocial African mole-rats, the naked mole-rat (Heterocephalus glaber) and the Damaraland mole-rat (Cryptomys damarensis)

Reproductive tracts and spermatozoa from reproductively active and reproductively suppressed non-breeding males from two species of eusocial African mole-rats Cryptomys damarensis and Heterocephalus glaber were examined. In two captive colonies of Heterocephalus glaber, reproductive tracts from seven non-breeding males removed from their colonies, and housed singly for 5-6 weeks to cause reproductive activation, were compared with reproductive tracts from seven non-breeding males. The body weight of the separated, reproductively active males increased significantly (P < 0.01), and the mean testis weights relative to body weight of the reproductively active males were significantly larger (P < 0.05) than those of non-breeding males. The number of spermatozoa, in one half of the reproductive tract, was higher in active males than in non-breeding males (mean +/- SEM: 8.59 x 10(6) +/- 2.69 x 10(6) versus 1.78 x 10(6) +/- 1.43 x 10(6), respectively; P < 0.05). In addition, six of the seven reproductively active males, but only two of seven non-breeding males, had motile spermatozoa. A total of 28 wild Cryptomys damarensis from two colonies were examined in the field. The testis weights relative to body weight of breeding males (n = 7) were higher than those of non-breeding males (n = 19; P < 0.01), but the number of spermatozoa did not differ significantly between the two groups (0.13 x 10(6) +/- 0.06 x 10(6), n = 7 versus 0.29 x 10(6) +/- 0.14 x 10(6), n = 21, respectively). Breeding and non-breeding males produced similar numbers of motile spermatozoa.(ABSTRACT TRUNCATED AT 250 WORDS)