Rapid Estimation on the Sperm Concentration in Boar Semen by Spectrophotometric Method

[ Objective] To establish the standardized spectrophotometric method to determine boar sperm concentration. [ Method ] The relation- ships between absorbance （A）, transmittance （T） and sperm concentration （C） of different wavelengths （450, 550, 650 nm） were compared. [ Result] The maximum sperm concentration detected by absorbance presented an upward trend with the increase of the wavelengths, 202 mitliorVml （450 nm）, 224 million/ml （550 nm） and 235 mUlion/ml （650 nm）, respectively, but the stability of repeated measurement was decreased. With the increase of sperm dilution times, the stability of repeated measurement of transmittance was reduced, and when dilution times were more than 10 times （450 nm）, 6 times （550 nm） and 4 times （650 nm）, differences appeared between the observed values of repeated measurement. [ Con- clusion] Wavelength at 450 nm was found to be the most sensitive and reliable, and sperm concentration presented cubic functional regression rela- tionship or power functional regression relationship with absorbance or transmittance, respectively. The regression equation for the standard curve at 450 nm was C400 = 0.48A3 - 0.76A2 ＋ 0.67A - 0.066 （ R = 0.951 ） and C400 = 1.657T -0.108. 8 （ R = 0.940）.

Semen analysis and sperm function testing

Despite controversy regarding the clinical value of semen analysis, male fertility investigation still relies on a standardized analysis of the semen parameters. This is especially true for infertility clinics in both developing and developed countries. Other optional tests or sophisticated technologies have not been widely applied. The current review addresses important changes in the analysis of semen as described in the new World Health Organization （WHO） manual for semen analysis. The most important change in the manual is the use of evidence-based publications as references to determine cutoff values for normality. Apart from the above mentioned changes, the initial evaluation and handling methods remain, in most instances, the same as in previous editions. Furthermore, the review evaluates the importance of quality control in andrology with emphasis on the evaluation of sperm morphology. WHO sperm morphology training programmes for Sub-Saharan countries were initiated at Tygerberg Hospital in 1995. The external qualitY control programme has ensured that the majority of participants have maintained their morphological reading skills acquired during initial training. This review reports on current sperm functional tests, such as the induced acrosome reaction, and sperm-zona pellucida binding assays, as well as the impact of sperm quality in terms of DNA integrity, and the relationship of sperm function tests to sperm morphology.

Predictors for partial suppression of spermatogenesis of hormonal male contraception

Aim： To analyze factors influencing the efficacy of hormonal suppression of spermatogenesis for male contraception. Methods： A nested case-control study was conducted, involving 43 subjects, who did not achieve azoospermia or severe oligozoospermia when given monthly injections of 500 mg testosterone undecanoate （TU）, defined as partial suppressors compared with 855 subjects who had suppressed spermatogenesis （complete suppressors）. Sperm density, serum testosterone, luteinizing hormone （LH） and follicle stimulating hormone （FSH） concentrations at the baseline and the suppression phase were compared between partial and complete suppressors. Polymorphisms of androgen receptor （AR） and three single nucleotide variants and their haplotypes of FSH receptor （FSHR） genes determined by polymerase chain reaction （PCR） and DNA sequencing technique were compared between 29 partial and 34 complete suppressors. Results： Baseline serum LH level was higher and serum LH as well as FSH level during the suppression phase was less suppressed in partial suppressors. Additionally, in a logistic regression analysis larger testis volume, higher serum FSH concentrations alone, or interaction of serum LH, FSH, testosterone and sperm concentrations were associated with degree of suppression. The distribution of polymorphisms of AR or FSH receptor genes did not differ between partial and complete suppressors. In cases with incomplete FSH suppression （FSH 〉 0.2 IU/L）, the chances of reaching azoospermia were 1.5 times higher in the subjects with more than 22 CAG triplet repeats. Conclusion： Partial suppression of spermatogenesis induced by 500 mg TU monthly injections is weakly influenced by hormonal and clinical features but not polymorphism in AR and FSHR genes.

Do Ureaplasma urealyticum infections in the genital tract affect semen quality?

Aim： To investigate the relationship between Ureaplasma urealyticum （UU） infection and semen quality. Methods： From 2001 to 2003, 346 eligible patients aged 20-45 years were invited from two hospitals in Shanghai, China, to participate in an investigation which included questionnaires about general and reproductive health, an external genital tract examination, UU culture and semen analysis. Multiple linear regression models were used to examine whether UU had a significant effect on semen quality after adjustment for confounding factors. Results： Findings suggested that UU infection was associated with higher semen viscosity and lower semen pH value. Sperm concentration was lower in UU positive subjects than that in UU negative subjects （54.04 × 10^6/mL vs.70.58 × 10^6/mL）. However, UU did not significantly affect other semen quality indexes. Conclusion： UU infection of the male genital tract could negatively influence semen quality.

Practical semen analysis： from A to Z

Accurate semen analysis is critical for decisions about patient care, as well as for studies addressing overall changes in semen quality, contraceptive efficacy and effects of toxicant exposure. The standardization of semen analysis is very difficult for many reasons, including the use of subjective techniques with no standards for comparison, poor technician training, problems with proficiency testing and a reluctance to change techniques. The World Health Organization （WHO） Semen handbook （2010） offers a vastly improved set of standardized procedures, all at a level of detail that will preclude most misinterpretations. However, there is a limit to what can be learned from words and pictures alone. A WHO- produced DVD that offers complete demonstrations of each technique along with quality assurance standards for motility, morphology and concentration assessments would enhance the effectiveness of the manual. However, neither the manual nor a DVD will help unless there is general acknowledgement of the critical need to standardize techniques and rigorously pursue quality control to ensure that laboratories actually perform techniques ＇according to WHO＇ instead of merely reporting that they have done so. Unless improvements are made, patient results will continue to be compromised and comparison between studies and laboratories will have limited merit.

A comparison of conventional and computer-assisted semen analysis.（CRISMAS software） using samples from 166 young Danish men

The aim of the present study was to compare assessments of sperm concentration and sperm motility analysed by conventional semen analysis with those obtained by computer-assisted semen analysis （CASA） （Copenhagen Rigshospitalet Image House Sperm Motility Analysis System （CRISMAS） 4.6 software） using semen samples from 166 young Danish men. The CRISMAS software identifies sperm concentration and classifies spermatozoa into three motility categories. To enable comparison of the two methods, the four motility stages obtained by conventional semen analysis were, based on their velocity classifications, divided into three stages, comparable to the three CRISMAS motility categories： rapidly progressive （A）, slowly progressive （B） and non-progressive （C＋ D）. Differences between the two methods were large for all investigated parameters （P〈0.001）. CRISMAS overestimated sperm concentration and the proportion of rapidly progressive spermatozoa and, consequently, underestimated the percentages of slowly progressive and non-progressive spermatozoa, compared to the conventional method. To investigate whether results drifted according to time of semen analysis, results were pooled into quarters according to date of semen analysis. CRISMAS motility results appeared more stable over time compared to the conventional analysis; however, neither method showed any trends. Apparently, CRISMAS CASA results and results from the conventional method were not comparable with respect to sperm concentration and motility analysis. This needs to be accounted for in clinics using this software and in studies of determinants of these semen characteristics.

Sperm origins and concentration do not impact the clinical outcomes in intracytoplasmic sperm injection cycles

In the present study, we evaluated the impact of sperm origins and concentration on the clinical outcomes of intracytoplasmic sperm injection （ICSI） cycles. A total of 1201 IC51 cycles were retrospectively analyzed for male azoospermia or oligozoospermia between January 2015 and December 2015 in the Peking University Third Hospital. Patients were divided into three groups （Group 1 vs Group 2/3; surgically extracted sperm vs ejaculated sperms）： Group 1 included 343 ICSI cycles and Group 2 analyzed 388 cycles on semen with sperm concentration 〈5 × 10^6 m1-1 （severe oligozoospermia group）. Group 3 included 470 cycles with sperm concentration between 5×10^6 m1-1 and 15 × 10^6 m1-1 （mild oligozoospermia group）. Fertilization rates, clinical pregnancy rates, and live birth rates were analyzed and compared among groups of different semen origins and concentrations on the oocyte retrieval day. Group 2 showed a lower fertilization rate than Group 3 （62.9%± 21.6% vs 66.8% ± 22.1%, P 〈 0.05）. There were no statistically significant differences in clinical pregnancy rate per transfer （51.3%, 46.7%, and 50.0%, respectively）, live birth rate per transfer （44.4%, 40.9%, and 41.4%, respectively）, accumulative live birth rate （58.3%, 51.0%, and 52.1%, respectively）, twin birth rate （18.4%, 10.6%, and 12.6%, respectively）, and birth defects rate （0, 0.3%, and 0.2%, respectively） among three groups. The results of this study indicated that sperm origins and concentration do not imDact the clinical outcomes in ICSI cycles.

Single semen analysis as a predictor of semen quality： clinical and epidemiological implications

It is generally thought that a single ejaculate is a bad predictor of semen quality of a subject, because of significant intra-individual variation. Therefore, we investigated the degree to which the results of a first semen analysis differ from that of a second analysis among men from a how the two different semen results mirrored the overall general population in Norway. In addition, we analysed semen quality assessment. A total of 199 volunteers participated in the study and delivered two semen samples with an interval of 6 months. The semen parameters were determined according to the World Health Organization （WHO） 1999 guidelines, which were also used to determine whether semen quality was normal or abnormal. In addition, the DNA fragmentation index （DFI） was determined using the Sperm Chromatin Structure Assay. The two samples from each individual were very similar with regard to standard semen parameters and DFI （rs： 0.67-0.72）, and there were no significant systematic differences between the two samples. The result of the first sample （normal/abnormal） was highly predictive of the overall conclusion based on the two samples （sperm concentration： in 93% of the cases （95% confidence interval [CI]： 89%-96%）; sperm motility： in 85% of the cases （95% CI： 79%-89%）; overall semen quality： in 85% of the cases （95% CI： 80%-90%）. In epidemiological studies, one ejaculate is a sufficient indicator of semen quality in a group of subjects. In a clinical situation, when the question is whether the semen quality is normal or not, the first ejaculate will, in at least 85% of cases, give a correct overall conclusion.

Viability of Extended Goat Semen Stored at Refrigerated Condition

Due to the small volume of goat semen ejaculate, just a few doses of goat semen were produced when the sperm concentration is 100 × 10^/mL. The study was aimed to determine the viability of extended goat semen at refrigerated condition at 5 ℃ using varying sperm concentrations and evaluated if sperm concentration lower than 100 × 10^6/mL would affect the motility, viability and sperm morphology at refrigerated condition. Using an artificial vagina, ejaculated goat semen was collected from goat semen donor aged 1.5 year. Physical evaluation of the collected semen showed an average volume of 0.54 mL, mean pH of 6.8 and a milky white color with thick consistency indicative of high concentration. Fresh goat semen had an initial average of 76% with an average initial sperm concentration of 128 × 10^/mL. The semen was divided into four treatments： sperm concentration of 100× 10^/mL, 75 × 10^/mL, 50 × 10^/mL and 25 × 10^/mL, and were stored at 5 ℃ for a period of 10 d. The semen evaluation was performed for each of the four treatments every other day. Results showed that the sperm concentration of spermatozoa affected the duration of storage based on the sperm motility percentage, viability and morphology of spermatozoa. The extended goat semen with sperm concentration of 25 and 50 million sperm/mL is optimal for storage within 6 d that gave satisfactory percentage motile, viable and morphologically normal spermatozoa.

Current treatment for male infertility:an umbrella review of systematic reviews and meta-analyses

This umbrella review aimed to summarize and provide a general evaluation of the effectiveness of current treatments for male infertility and assess the quality of evidence and possible biases.An umbrella review of systematic reviews and meta-analyses available in PubMed,Web of Science,and Scopus,covering studies published up to October 2023,was conducted.Sperm concentration,morphology,and motility were used as endpoints to evaluate the effectiveness of the treatments.Of 2998 studies,18 published meta-analyses were extracted,yielding 90 summary effects on sperm concentration(n=36),sperm morphology(n=26),and sperm motility(n=28)on 28 interventions.None of the meta-analyses were classified as having low methodological quality,whereas 12(66.7%)and 6(33.3%)had high and moderate quality,respectively.Of the 90 summary effects,none were rated high-evidence quality,whereas 53.3%(n=48),25.6%(n=23),and 21.1%(n=19)were rated moderate,low,and very low,respectively.Significant improvements in sperm concentration,morphology,and motility were observed with pharmacological interventions(N-acetyl-cysteine,antioxidant therapy,aromatase inhibitors,selective estrogen receptor modulators,hormones,supplements,and alpha-lipoic acid)and nonpharmacological interventions(varicocele repair and redo varicocelectomy).In addition,vitamin supplementation had no significant positive effects on sperm concentration,motility,or morphology.Treatments for male infertility are increasingly diverse;however,the current evidence is poor because of the limited number of patients.Further well-designed studies on single treatment and high-quality meta-analysis of intertreatment comparisons are recommended.