Sperm Functional Parameters Research Articles

Effect of exogenous DNA on bovine sperm functionality using the sperm mediated gene transfer (SMGT) technique

Sperm mediated gene transfer (SMGT) could provide the opportunity to carry out transgenesis on a mass scale using spermatozoa as vectors for exogenous DNA. However, the efficiency of sperm-mediated DNA transfer is still questionable, and the mode of transmission to the egg has not yet been well understood. Our aim was to investigate the capacity of bovine spermatozoa to carry exogenous DNA and its relationship to sperm functionality. We studied these parameters using flow cytometry to measure viability (necrosis and apoptosis) and capacitation status, computer-assisted semen analysis (CASA) to measure motility parameters and in vitro fertilization (IVF) to assess fertilizing capacity. Furthermore, we studied the effect of capacitation status on interaction with exogenous DNA, and the role of heparin supplementation in this process. Bull spermatozoa showed a high capacity to bind DNA quickly and reached a maximum after 30 min, with approximately half of the DNA-bound spermatozoa being viable. Incubation with exogenous DNA induced a decrease in sperm viability and motility and increased the proportion of apoptotic cells, but did not affect the cleavage rate in IVF assay. Heparin increased high-lipid disorder and the number of sperm with DNA bound (viable and dead). In conclusion, this study shows that live spermatozoa can bind exogenous DNA with a slight negative effect in some parameters of sperm function that in our opinion, would not drastically compromise fertility.

The Effect of Cyperus esculentus on Sperm Function Parameters in Prepubertal Mice as a Model for Human

The objective of this work was to study the effect of oral administration of Cyperus esculentus (CE) and its alcoholic extract on sperm function parameters in prepubertal mice as a model for human .The animals were divided into three groups each contains 6 animals .Group 1 was treated with 150 mg/ kg body weight /day of crude CE, group 2 was treated with same dose of alcohol extract of CE and group 3 regarded as control throughout six weeks period. The results showed a significant (p> 0.05) increase in the mean of sperm concentration ,sperm motility percent and progressive sperm motility between treated groups and control . There was no differences among groups in the mean of sperm normal morphology and sperm viability . No significant differences was recorded in the mean of body weight among groups throughout the study. The results revealed that the administration of Cyperus esculentus may enhance certain sperm characters in prepubertal mice without affecting body weight.

123. THE EFFECT OF PATERNAL DIET INDUCED OBESITY ON SPERM CAPACITATION, ACROSOME REACTION, BINDING AND FERTILISATION IN MOUSE MODEL

While the effects of obesity on male fertility are emerging, the direct effects on sperm function are less clear. The aim of this study was to determine the effects of diet-induced obesity on sperm capacitation, acrosome reaction, oocyte binding and fertilisation. C57/Bl6 male mice (n = 12) were randomly allocated to two groups; group 1 received a control diet (6% fat) while group 2 received a high fat diet (HFD, 22% fat) for up to 14 weeks. Mice were sacrificed and spermatozoa obtained. Capacitation and acrosome reaction were measured using Arachis hypogaea (peanut) agglutinin. Sperm binding to oocytes was assessed by co-incubation of sperm with superovulated cumulus-enclosed oocytes for 4 hrs. Fertilisation rates were expressed as the percentage of oocytes with 2 pronuclei from the total number inseminated 6 hrs post insemination. The percentage of non-capacitated sperm in males fed a high fat diet was significantly lower compared to males fed a control diet (12.3% vs 21.1%; P < 0.01). The percentage of acrosome reacted sperm did not differ between the groups. Following 4 hrs of co-incubation with cumulus-enclosed oocytes, the number of sperm bound to each oocyte was significantly lower in the HFD group compared to controls (41.14±2.5 vs 58.39±2.4; P < 0.01). Moreover, the percentage of fertilized oocytes was significantly lower in the HFD group compared to controls (25.9% vs 43.9; P < 0.01). This study demonstrates that males fed a HFD to induce obesity have impaired spermatozoa as evidenced by lower levels of capacitation and reduced ability to bind and fertilise an oocyte. These data therefore provide direct evidence that the metabolic health of the male can have a significant impact on sperm function parameters that are associated with infertility.

Chromomycin A3 staining, sperm chromatin structure assay and hyaluronic acid binding assay as predictors for assisted reproductive outcome

Functional sperm tests such as the sperm chromatin structure assay (SCSA), chromomycin A3 staining (CMA(3)) and hyaluronic acid binding assay (HBA) have been suggested as predictive tests of fertility in vitro. This study aimed to define the clinical role of these functional parameters in assisted reproduction in a prospective cohort study. Conventional sperm diagnosis (motility, morphology and concentration) as well as SCSA, CMA(3) and HBA tests were performed on 205 semen samples [74 IVF, 94 ICSI and 37 combined IVF/intracytoplasmic sperm injection (ICSI)]. Main outcome parameters were fertilization rate, clinical pregnancy rate and take-home baby rate. The study showed that each of the three functional sperm tests was related to one or more conventional and one or more functional sperm tests, indicating that spermatozoa from patients with abnormal conventional semen parameters have a higher likelihood for multiple functional abnormalities. Only SCSA and CMA(3) staining were shown to have a limited predictive value when IVF or combined IVF/ICSI was applied. The proposed threshold value of <or=15% for predicting good fertilization rates and obtaining a pregnancy in IVF could only be confirmed for percent HDS (high DNA stainability in SCSA). ICSI outcome was not influenced by any of the conventional or functional sperm parameters.

Improvement in buffalo ( Bubalus bubalis) spermatozoa functional parameters and fertility in vitro: Effect of insulin-like growth factor-I

The aim of the current study was to assess the effect of insulin-like growth factor-I (IGF-I; 100 ng/mL) on buffalo ( Bubalus Bubalis) sperm functional parameters related to in vitro fertilization. The acrosin activity (the mean diameter of halo formation in micrometers) was significantly higher in the IGF-I group (14.17 ± 1.51) compared with that in the control group (9.50 ± 0.36) at 2 h incubation. The mitochondrial membrane potential (per cent) was significantly higher in the IGF-I group compared with that in the control group at 30 min (33.27 ± 2.62 vs. 26.71 ± 1.02), 60 min (24.24 ± 3.45 vs. 18.77 ± 2.09), and 90 min (22.86 ± 3.02 vs. 16.92 ± 1.24) incubation. The percentage of spermatozoa positive for sperm nuclear chromatin decondensation (NCD) differed significantly between the groups at 90 and 120 min incubation. The comet length was significantly lower in the IGF-I group compared with that in the control group at 2 h incubation. The percentage of fragmented DNA in the tail did not differ significantly between the groups at 2 h incubation. The percentage of acrosomal-reacted spermatozoa did not differ significantly between the IGF-I and the control groups at 4 h (41.12 ± 6.44 vs. 43.53 ± 5.05) incubation. The cleavage rate (per cent) was significantly higher in the IGF-I–treated group (56.73 ± 3.70) compared with that in the control group (44.85 ± 2.15). The current study suggests that the addition of IGF-I prevents deterioration of sperm functional parameters and fertility.

Adverse effects of cadmium exposure on mouse sperm

The effects of cadmium chloride exposure on sperm functional parameters were evaluated on eight-week-old ICR-CD1 male mice administered with a single s.c. injection of 1, 2 and 3 mg CdCl 2/kg bw. Groups of animals treated with each dose, as well as their respective controls, were sacrificed after 24 h to detect short-term (acute) effects and after 35 days. Sperm cells were collected from the epididymis and several parameters of sperm quality and function were evaluated, namely density, morphology, motility, viability, mitochondrial function, acrosome integrity, together with DNA fragmentation assessed by the TUNEL assay. The short-term effects of cadmium chloride resulted in an increased fraction of sperm with abnormal morphology, premature acrosome reaction and reduced motility. Late term effects (after 35 days) included a drastic reduction of sperm cell numbers and sperm motility. An increase in DNA fragmentation was also detected.

Freezability prediction of boar ejaculates assessed by functional sperm parameters and sperm proteins

The objective of this work was to look for useful predictive indicators of the potentially “good” or “poor” ability of a boar ejaculate to sustain cryopreservation by assessing both the conventional sperm quality parameters (Study 1) and the immunolabeling of three proteins involved in the physiology of the sperm cell: GLUT3, HSP90AA1 and Cu/ZnSOD (Study 2). Study 1 was carried out in three different steps during the cryopreservation process of the sperm-rich fraction of 29 Piétrain boar ejaculates (17 °C, 5 °C, and 240 min postthaw). These ejaculates were clustered based on sperm quality parameters analyzed at 240 min postthaw, obtaining 16 good freezability ejaculates (GFEs) and 13 poor freezability ejaculates (PFEs). The sperm linearity (LIN) and the straightforward (STR) indexes at 5 °C showed higher hyperactivated movement in the PFEs than in the GFEs, which suggests that analyzing these sperm kinematic parameters could be a useful tool for predicting the potential freezability of an ejaculate. This statement was demonstrated by grouping the 29 ejaculates into two clusters (A and B) based on LIN and STR values assessed after 30 min at 5 °C, which resulted in around 72% of coincidence with the GFE and PFE groups. Study 2, performed at 17 °C and 240 min postthaw, revealed no differences between GFEs and PFEs in the immunolabeling of the three proteins within a same step, in terms of location and reactivity, although reactivity was generally weaker at 240 min postthaw in both groups. Additional studies on Western blot are currently being carried out with the objective to quantify the expression of the three proteins in GFEs and PFEs in the three steps of the cryopreservation process.

Methyl-parathion decreases sperm function and fertilization capacity after targeting spermatocytes and maturing spermatozoa

Paternal germline exposure to organophosphorous pesticides (OP) has been associated with reproductive failures and adverse effects in the offspring. Methyl-parathion (Me-Pa), a worldwide-used OP, has reproductive adverse effects and is genotoxic to sperm, possibly via oxidative damage. This study investigated the stages of spermatogenesis susceptible to be targeted by Me-Pa exposure that impact on spermatozoa function and their ability to fertilize. Male mice were exposed to Me-Pa (20 mg/kg bw, i.p.) and spermatozoa from epididymis-vas deferens were collected at 7 or 28 days post-treatment (dpt) to assess the effects on maturing spermatozoa and spermatocytes, respectively. Spermatozoa were examined for DNA damage by nick translation (NT-positive cells) and SCSA (%DFI), lipoperoxidation (LPO) by malondialdehyde production, sperm function by spontaneous- and induced-acrosome reactions (AR), mitochondrial membrane potential (MMP) by using the JC-1 fluorochrome, and fertilization ability by an in vitro assay and in vivo mating. Alterations on DNA integrity (%DFI and NT-positive cells) in spermatozoa collected at 7 and 28 dpt, and decreases in sperm quality and induced-AR were observed; reduced MMP and LPO were observed at 7 dpt only. Negative correlations between LPO and sperm alterations were found. Altered sperm functional parameters evaluated either in vitro or in vivo were associated with reduced fertilization rates at both times. These results show that Me-Pa exposure of maturing spermatozoa and spermatocytes affects many sperm functional parameters that result in a decreased fertilizing capacity. Oxidative stress seems to be a likely mechanism of the detrimental effects of Me-Pa exposure in male germ cells.

Scavenging effect of N-acetyl-L-cysteine against reactive oxygen species in human semen: a possible therapeutic modality for male factor infertility?

A new approach to reduce the level of reactive oxygen species (ROS) in human semen by using N-acetyl-L-cysteine (NAC) was evaluated. Semen samples were incubated with or without NAC (1.0 mg ml-1) at room temperature. The chemiluminescent signal of the oxidation of luminol was detected by means of an MTP reader after 0, 20, 40, 60 and 120 min, respectively, using 200 microM luminol. In addition, the dose-dependent action of NAC (0.1, 1.0 and 5.0 mg ml-1) and the influence of NAC on functional sperm parameters (motility and acrosome reaction) were studied. ROS levels decreased significantly after 20 min incubation with NAC. This reduction was greater in the high ROS group (> 30000 counts/10(7) viable sperm at t = 0) than in the low ROS group (< 30000). In addition, a marked dose-dependence of NAC was observed. Concerning sperm function, total sperm motility improved after incubation with NAC, but no significant change was observed with respect to the acrosome reaction. NAC (at concentrations of 1.0 mg ml-1) significantly reduced ROS in human semen and showed the possibility of improving impaired sperm function. After further testing NAC might be useful for the treatment of male infertility patients.

Pre-freeze bull sperm head morphometry related to post-thaw fertility

Artificial insemination using cryopreserved semen is a common management tool of the contemporary livestock producer. The ability to determine post-thaw fertility from pre-freeze sperm function parameters would be of major benefit to producers. In this study computer-aided sperm head morphometry was used to determine whether there is any association between pre-freeze bull sperm head measurements and post-thaw non-return to oestrus rates. Sixteen commercial artificial insemination bulls were used for this study. Of the 16 bulls, eight had ≥69% non-return to oestrus rates while the remaining eight bulls had <69% non-return to oestrus rates, based on artificial insemination of cryopreserved semen from a minimum of 748 inseminations per bull. Microscope slides of extended or cryopreserved and thawed semen were prepared and stained by the MGZIN procedure. The morphometric dimensions for length, width, width/length, area and perimeter for a minimum of 200 sperm heads were analyzed from each slide by ASMA and the mean measurements and intra-analysis coefficients of variation (CV) recorded. The post-thaw non-return to oestrus rates for all bulls were correlated with pre-freeze measurements of width ( r = 0.53, P < 0.05), and the change in width/length after cryopreservation ( r = 0.61, P < 0.05). There were no significant differences in the mean pre-freeze or post-thaw sperm head measurements between the two groups. However, the mean post-thaw change in width/length was significantly ( P = 0.003) different between the groups. The mean post-thaw intra-analysis variability for width was significantly lower ( P = 0.03) in bulls with non-return to oestrus rates (NRR) ≥ 69%. Overall, the data suggests that pre-freeze and post-thaw bull sperm head morphometry measurements of individual bulls may have minimal predictive value of post-thaw fertility as determined by non-return to oestrus rates between groups of bulls with limited variance in non-return to oestrus rates. The clinical relationship of sperm head morphometry and fertility in bulls with a greater, clinically significant range of non-return to oestrus, remains to be studied.

Poor sperm quality negatively impacts embryo development and reproductive outcome in donor oocyte assisted reproductive technology cycles

OBJECTIVE: Oocyte donation offers a powerful tool in the study of sperm quality, as oocyte and endometrial characteristics can be controlled and are comparable among the recipients. Since the introduction of intracytoplasmic sperm injection (ICSI), little attention is given to specific parameters of sperm function and their impact on IVF outcome. Our aim is to determine whether semen parameters influences embryo development and reproductive potential, and if they could be regarded as a prognostic factor in donor assisted reproductive technology cycles. DESIGN: Retrospective cohort analysis. MATERIALS AND METHODS: Three hundred and thirty two fresh ICSI cycles using donor oocytes were analyzed. ICSI was performed routinely to avoid inmature oocytes being given to the recipients. Only cycles in which >3 embryos were yielded and at least two Day3 good quality embryos were available for ET were included. Male partners were classified as normal (N), low-moderate (LM) or severe (S) male factor infertility according to the following sperm parameters: 1) Percent normal morphology by strict Kruger criteria: (S) <4%, (LM) 4-14%, (N)>14%, 2) Sperm concentration: (S) <10million/ml, (LM) 10- 20 million/ml, and (N) > 20million/ml, 3) Motility (L) <30%, (LM) 30-50%, (N) >50%, and 4) Post-preparation total motile count: (S) < 5 million, (LM) 5-10 million, and (N)>10 million sperm. Male factor infertility was correlated to a variety of outcome measures: embryo quality (day-3 cell cleavage, and fragmentation), fertilization, clinical pregnancy rate (CPR) and pregnancy loss. Statistical analysis was performed with chi2 for categorical data and t test for continuous parametric data. RESULTS: The number of fertilized oocytes and the number of day 3 embryos >7 cells was similar among the normal and low-moderate factor infertility group. However, the number of good quality embryos available for ET were significantly lower in the severe male factor infertility group (P<0.005). Implantation and clinical pregnancy rates were significantly decreased in this group when compared to male partners with normal or low-moderate semen parameters: Normal CPR 52,05% (76/146), Low-Moderate CPR 40,91%, Severe CPR 29,63% (16/54) (p<0.05). CONCLUSIONS: Poor sperm quality was negatively associated with reproductive outcomes in oocyte donation cycles suggesting a paternal influence on embryo development in which ICSI may not be able to select the sperm of highest quality for oocyte injection.

Direct contact between boar spermatozoa and porcine oviductal epithelial cell (OEC) cultures is needed for optimal sperm survival in vitro

Oviductal epithelial cell (OEC) co-culture prolongs sperm viability and motility in vitro in a number of species including humans and horses. This study has sought to determine the effects of homologous OEC co-culture on boar sperm function. To determine whether the effects on spermatozoa were specifically caused by co-culture with or by OEC secretions, or by both factors together, a number of co-culture and cell-conditioned medium (CM) experiments were conducted. Firstly, Percoll-washed spermatozoa were co-cultured with OECs and pig kidney epithelial (LLC-PK1) cells, and in medium without cells. Secondly, Percoll-washed spermatozoa were incubated with CM derived from both OECs and LLC-PK1 cells and in unconditioned medium. A number of sperm function parameters were assessed after 5, 30, 60, 90, 120, and 180 min, and 24 h of co-culturing or incubation with CM. Of all the sperm function parameters investigated, the percentage (%) viability data yielded the most interesting results. OECs (mean ± S.E.M.; 31.2 ± 1.10) were better than LLC-PK1 cells (24.3 ± 0.93) at prolonging the viability of unbound spermatozoa after 24 h of co-culturing ( P < 0.05). Also after 24 h, the viability of spermatozoa bound to the OECs (77.6 ± 1.83) was significantly higher than in the case of the LLC-PK1 cells (53.5 ± 1.43; P < 0.001). Other sperm function parameters, e.g., capacitation and motility, were also influenced by OEC co-culturing and incubation with CM, although to a lesser degree. In conclusion, porcine homologous OEC co-culture and CM incubation specifically affect sperm function. However, we propose that it is OEC co-culturing, rather than OEC-CM, that has the greater influence.

Lead chloride affects sperm motility and acrosome reaction in mice

Lead is highly toxic and persistent in the environment and, thus, a major concern for public health. In this study, the effects of lead chloride (PbCl2) on mouse epididymal sperm were evaluated. Male mice were subcutaneously injected with 74 and 100 mg PbCl2/kg body weight for four consecutive days. Sperm was collected from the epididymis and several parameters of sperm function, such as sperm density, motility, viability, mitochondrial function, acrosome integrity and morphology, were evaluated. Furthermore, DNA fragmentation was assessed by the terminal deoxylnucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick-end labelling (TUNEL) assay and chromatin integrity was evaluated by sperm chromatin structure assay (SCSA). In order to assess direct effects on existing sperm population, we sacrificed one group for each condition at day 5. The effects of lead upon one entire spermatogenic cycle were evaluated on day 35. Both lead concentrations used in this work affected sperm motility, although no significant differences were observed in sperm viability, mitochondrial function and DNA/chromatin integrity. However, a decrease in the percentage of intact acrosomes was also observed, mirroring a lead-induced premature acrosome reaction. Thus, the results obtained indicate that, together with impaired motility, the effect of lead toxicity on acrosome integrity, leading to premature reaction, may compromise the ability of sperm to fertilize the oocyte.

Is abnormal sperm function an indicator among couples with recurrent pregnancy loss?

To determine whether or not sperm function parameters are altered in male partners of couples with a history of idiopathic recurrent pregnancy loss (RPL). In comparison with proven fertile volunteers, sperm function parameters like hypo-osmotic swelling (HOS), acrosomal status (AS), and nuclear chromatin decondensation (NCD) were assessed in vitro from male partners of couples with a history of idiopathic RPL. Infertility clinic and andrology laboratory at National Institute of Health and Family Welfare. Male partners of couples with a history of idiopathic RPL and proven fertile male volunteers (control). Standard semen analysis, assessment of sperm morphology, and sperm function with tests such as HOS, AS, and NCD. Sperm paameters, such as HOS, AS, and NCD, were assessed in semen samples from RPL in comparison with the proven fertile control group. Semen samples from the idiopathic RPL group showed below normal test scores in 57.1% of the cases for all three sperm parameters. The highest aberration (83% of cases) in sperm attributes was observed in NCD, followed by AS (45.7%) and HOS (42.9%). In contrast, abnormality in sperm morphology was limited to 5.7% of the cases. Subnormal sperm function is directly proportional with subnormal sperm motility (<50%) in 23% of the cases. Even in semen samples with normal sperm motility, sperm function scores were below normal in 31.4% of the RPL group. Reduction in test scores of sperm function, like HOS, AS, and NCD, in male partners of couples with idiopathic RPL suggests that sperms with altered or lowered functional competencies, if they fertilize the oocytes, may lead to the development of an unsustainable embryo resulting in early pregnancy loss. Normal sperm motility does not always ensure normal sperm function scores.

Moderate Seasonality in Testis Function of Domestic Cat

Adult male domestic cats are known to produce sperm throughout the year, although sexual activity is influenced by geographical location. In the northern hemisphere, feral domestic cats reproduce usually between January and July. Thus, seasonality in testicular activity might be suggested. The aim of the present study was to investigate gametogene and endocrine activity of cat testis throughout the entire year. Testes and epididymides (n = 10-12 per month) were collected after castration. Spermatogenesis was quantified by assessment of testicular sperm per testis and by flow cytometric analysis of the cells with different DNA content. Sperm from cauda epididymis were evaluated according to motility and morphological integrity. Testicular testosterone concentration was determined by enzyme immunoassay. Testis mass and sperm production varied moderately throughout the year. Significant seasonal variations were observed in the proportion of cells in the G2/M phase of cell cycle (p = 0.004) and the meiotic transformation (ratio of haploid : tetraploid cells; p = 0.021). Changes in testicular testosterone concentration were more pronounced and showed periods with high (spring) and significantly reduced testosterone levels (autumn). A marked seasonal alteration (p < 0.001) with a peak in March was assessed in the percentage of progressively motile sperm. The proportion of morphological intact sperm was also significantly higher in spring compared with winter time (p < 0.001). In conclusion, the study suggests moderate seasonal changes in quantity of sperm, more pronounced annual variation in hormone production and a distinct seasonal influence on functional sperm parameters in domestic cat.

Effect of breed and sperm concentration on the changes in structural, functional and motility parameters of ram-lamb spermatozoa during storage at 4 °C

The objectives of this study were (1) to determine the changes in structural, functional and motility parameters of ram-lamb semen stored at two different concentrations at 4 °C for 8 days in egg-yolk based extender and (2) to determine the effect of breed of ram-lambs on the changes in structural, functional and motility parameters of ram-lamb semen from different breeds stored at two different concentrations at 4 °C for 8 days in egg-yolk based extender. Two different concentrations suitable for laparoscopic and cervical insemination were employed in this experiment. A total of 14 ram-lambs (Polled Dorset—5, Suffolk—5, Katahdin—4) with satisfactory breeding potential were selected. Semen samples were collected by electro-ejaculation. Semen samples were extended to 50 and 200 million sperm per ml with a commercial egg yolk based extender (Triladyl ®, Minitube of America, Verona, WI, USA) at room temperature and were stored at 4 °C. The sperm DNA fragmentation index (DFI), percentages of high mitochondrial membrane potential (hMMP) and plasma membrane integrity (PMI) were assessed using flow cytometry as part of structural and functional parameters on Days 0, 1, 4, 6, and 8. A computer assisted sperm analyser (HTM-IVOS, Version 10.8, Hamilton Thorne Research, Beverly, MA, USA) was used to assess the sperm motility parameters on Days 0, 1, 4, 6, and 8. PROC MIXED procedure was used to determine the effect of days of storage, concentration and breed. The concentration and days of storage significantly affected the sperm structural, functional and motility parameters ( P < 0.0001). Significant concentration × days of storage interaction was found for all structural and functional parameters. There was a significant concentration × days of storage interaction for average path velocity, curvilinear velocity, straightness and linearity. Overall changes in the sperm structural, functional and sperm motility parameters over the storage period were less dramatic in the 200 × 10 6 ml −1 concentration when compared to 50 × 10 6 ml −1 concentration. The hMMP and total progressive motility were influenced by breed. In conclusion, the quality of structural, functional and motility parameters declined as days of storage were increased and the magnitude of changes in the parameters was less dramatic at the higher concentration.

P-627: Are routine semen parameters predictive of IVF outcome? A reassessment in the ICSI era using IVF and split sibling oocyte IVF-ICSI cohorts

Much attention has been focused on ovarian stimulation and its impact on pregnancy outcome. Particularly since the introduction of ICSI, little attention is given to specific parameters of sperm function and their impact on IVF outcome. Given the lack of good data regarding the sperm’s role in pregnancy outcome, we critically investigated all routine measures of sperm obtained in the standard andrology lab. In addition to low sperm strict morphology (<4% normal forms), we hypothesized that there may be additional sperm-related variables that can further define the need to proceed to ICSI during IVF.

Semen quality and sperm functional parameters in fertile Indian men

The reports of a decline in the reproductive health of men worldwide prompted the present study to be undertaken so that baseline semen parameters could be established in Indian men. Semen profile and sperm function parameters were evaluated in 368 Indian men of proven fertility, settled in Delhi. The results of the study were compared with available published information from Indian men. The mean sperm concentration and semen volumes were 68.22 +/- 15.14 x 10(6) ml(-1) and 3.20 +/- 0.94 ml, respectively. Rapid, linear progressive motility and sluggish linear motility were 40.95 +/- 9.15% and 24.95 +/- 7.01%, respectively. A comparison of the results of the present study with earlier published data did not support the contention of a decrease in the semen quality in Indian men.

Influence of cigarette smoking on spermatozoa via seminal plasma

Numerous investigations have been conducted on the relationship between cigarette smoking and male infertility, however, the exact molecular mechanisms are not well understood in most of the cases. Few studies have indicated the direct effect of seminal plasma (SP) [in different dilutions with phosphate buffer solution (PBS)] from smokers (SM) on the sperm functional parameters from nonsmokers (non-SM). The aim of this study was to provide evidence that cigarette smoking affects male fertility via altering the sperm quality. Our results indicated that exposure of spermatozoa from the non-SM to the SP from the SM yielded a significant reduction in the sperm motility and acrosome reaction and an elevation in the amount of malondialdehyde (MDA), in a certain time course. Exposure of spermatozoa from the SM to the SP from the non-SM or with PBS resulted in the nonsignificant improvement in the altered sperm functional parameters indicating removal of SM's SP and then subsequent reconstitution with physiological media could be of clinical significance in the various assisted reproductive technologies applied for SM. However, the detrimental effect of SM's SP on non-SM's spermatozoa was prominent. In addition, as spermatozoa in SM's SP are susceptible to peroxidative damages, men with such cells who wish to have children should especially benefit from quitting smoking.

Sperm function and assisted reproduction technology

The evaluation of different functional sperm parameters has become a tool in andrological diagnosis. These assays determine the sperm’s capability to fertilize an oocyte. It also appears that sperm functions and semen parameters are interrelated and interdependent. Therefore, the question arose whether a given laboratory test or a battery of tests can predict the outcome inin vitro fertilization (IVF). One-hundred and sixty-one patients who underwent an IVF treatment were selected from a database of 4178 patients who had been examined for male infertility 3 months before or after IVF. Sperm concentration, motility, acrosin activity, acrosome reaction, sperm morphology, maternal age, number of transferred embryos, embryo score, fertilization rate and pregnancy rate were determined. In addition, logistic regression models to describe fertilization rate and pregnancy were developed. All the parameters in the models were dichotomized and intra- and interindividual variability of the parameters were assessed. Although the sperm parameters showed good correlations with IVF when correlated separately, the only essential parameter in the multivariate model was morphology. The enormous intra- and interindividual variability of the values was striking. In conclusion, our data indicate that the andrological status at the end of the respective treatment does not necessarily represent the status at the time of IVF. Despite a relatively low correlation coefficient in the logistic regression model, it appears that among the parameters tested, the most reliable parameter to predict fertilization is normal sperm morphology.