• Animal Bioscience
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• v.34 no.8
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• pp.1253-1270
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• 2021

Assessment of male fertility is based on the evaluation of sperm. Semen evaluation measures various sperm quality parameters as fertility indicators. However, semen evaluation has limitations, and it requires the advancement and application of strict quality control methods to interpret the results. This article reviews the recent advances in evaluating various sperm-specific quality characteristics and methodologies, with the help of different assays to assess sperm-fertility status. Sperm evaluation methods that include conventional microscopic methods, computer-assisted sperm analyzers (CASA), and flow cytometric analysis, provide precise information related to sperm morphology and function. Moreover, profiling fertility-related biomarkers in sperm or seminal plasma can be helpful in predicting fertility. Identification of different sperm proteins and diagnosis of DNA damage has positively contributed to the existing pool of knowledge about sperm physiology and molecular anomalies associated with different infertility issues in males. Advances in methods and sperm-specific evaluation has subsequently resulted in a better understanding of sperm biology that has improved the diagnosis and clinical management of male factor infertility. Accurate sperm evaluation is of paramount importance in the application of artificial insemination and assisted reproductive technology. However, no single test can precisely determine fertility; the selection of an appropriate test or a set of tests and parameters is required to accurately determine the fertility of specific animal species. Therefore, a need to further calibrate the CASA and advance the gene expression tests is recommended for faster and field-level applications.

• Clinical and Experimental Reproductive Medicine
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• v.41 no.1
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• pp.9-14
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• 2014

Objective: The aim of this study was to evaluate the efficiency of the intracytoplasmic morphologically selected sperm injection (IMSI) technique compared with conventional ICSI and previous ICSI attempts in oligo-astheno-teratozoospermia (OAT) patients. Methods: The sperms were selected under high magnification ($6,600{\times}$) and used to induce fertilization in previous ICSI patients by IMSI. These results were compared with previous conventional ICSI cycles in patients with OAT infertility. Results: These results demonstrated no significant difference in the fertilization rate between IMSI and previous ICSI cycles (67.7% vs. 65.0%). However, the pregnancy and implantation rates with IMSI were significantly higher than those of the ICSI cycles (33.3% vs. 12.5% and 14.6% vs. 5.4%, respectively; p<0.05). The miscarriage rate among pregnant patients (18.2% vs. 37.5%) showed no statistically significant difference between groups. Conclusion: Compared to conventional ICSI, this study found that IMSI increased the IVF-ET success rates in patients with OAT.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.127-133
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• 2008

The objective of this study was two folds: to investigate the relationship between paternal identification rate and sperm quality parameters such as motility and sperm chromatin structure assay after heterospermic insemination; to see if mutual complement between tests and development of useful technique to enhance the fertility in artificial insemination. In individual boar's fertilizing ability, 3 high fertility boars showed significantly high fertility (p<0.05) compared to 3 low fertility boars, but there was no difference in litter size between two groups. Sperm motility test in pooled and individual semen using computer assisted sperm analysis (CASA) revealed that no significant difference among boars. The high fertile boar showed tendency of low %Red (High red fluorescence/green+red fluorescence) in sperm chromatin structure assay (SCSA) but paternal identification rate from piglets did not differ after heterospermic insemination. The correlation coefficient between individual or pooled semen function test and farrowing rates were well correlated as follows: %Red with litter size (r= - 0.53, p=0.03); %Red with paternal identification rates (r=-0.51, p=0.03); paternal identification rates with litter size (r=0.57, p=0.02). These results indicate that sperm chromatin structure assay and sperm quality parameter test in pooled semen are useful method to predict and evaluate the fertilizing capacity after heterospermic insemination in boars.

• Animal Bioscience
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• v.36 no.3
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• pp.385-403
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• 2023

Male fertility is affected by multiple endogenous stressors, including reactive oxygen species (ROS), which greatly deteriorate the fertility. However, physiological levels of ROS are required by sperm for the proper accomplishment of different cellular functions including proliferation, maturation, capacitation, acrosomal reaction, and fertilization. Excessive ROS production creates an imbalance between ROS production and neutralization resulting in oxidative stress (OS). OS causes male infertility by impairing sperm functions including reduced motility, deoxyribonucleic acid damage, morphological defects, and enhanced apoptosis. Several in-vivo and in-vitro studies have reported improvement in quality-related parameters of sperm following the use of different natural and synthetic antioxidants. In this review, we focus on the causes of OS, ROS production sources, mechanisms responsible for sperm damage, and the role of antioxidants in preserving sperm fertility.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.1
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• pp.22-29
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• 2015

Objective: Sperm must be properly prepared in in vitro fertilization (IVF)-embryo transfer (ET) programs in order to control the fertilization rate and ensure that embryos are of high quality and have appropriate developmental abilities. The objective of this study was to determine the most optimal sperm preparation method for IVF. Methods: Patients less than 40 years of age who participated in a fresh IVF-ET cycle from November 2012 to March 2013 were included in this study. Poor responders with less than three mature oocytes were excluded. Ham's F-10 medium or sperm-washing medium (SWM) was used in combination with the density-gradient centrifugation/swim-up (DGC-SUP) or SUP methods for sperm preparation. A total of 429 fresh IVF-ET cycles were grouped according to the media and methods used for sperm preparation and retrospectively analyzed (DGC-SUP/Ham's F-10, n=82; DGC-SUP/SWM, n=43; SUP/Ham's F-10, n=181; SUP/SWM, n=123). Results: There were no significant differences among these four groups with respect to the mean age of the female partners, duration of infertility, number of previous IVF cycles, and retrieved oocytes. We determined that both the DGC-SUP and SUP methods for sperm preparation from whole semen, using either Ham's F-10 or SWM media, result in comparable clinical outcomes, including fertilization and pregnancy rates. Conclusion: We suggest that both media and both methods for sperm preparation can be used for selecting high-quality sperm for assistive reproductive technology programs.

• Animal Bioscience
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• v.36 no.11
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• pp.1647-1654
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• 2023

Objective: Aging roosters typically exhibit subfertility with decreasing semen quality, furthermore Thai native roosters reared in rural areas are raised for a longer duration than their usual lifespan. The present study therefore aimed to assess the effect of selenium supplementation as an antioxidative substance in diets to improve the semen cryopreservation of aged roosters. Methods: Semen samples were collected from young (n = 20) and aged (n = 20) Thai native roosters (Pradu Hang Dum) at 36 and 105 weeks of age when starting the experiment, respectively. They were fed diets either non-supplemented or supplemented with selenium (0.75 ppm). Fresh semen quality and lipid peroxidation of fresh semen was evaluated before cryopreservation using the traditional liquid nitrogen vapor method. Post-thaw sperm quality and fertility potential were determined. Results: Advancing age is unrelated to decreasing fresh semen quality (p>0.05). However, lipid peroxidation in rooster semen depended on age, and the malondialdehyde (MDA) concentration increased in aged roosters (p<0.05). Selenium supplementation in diets significantly decreased the MDA concentration and increased the sperm concentration (p<0.05). In contrast, cryopreserved semen was affected by advancing rooster age, and selenium influenced sperm quality (p<0.05). Younger roosters had higher post-thaw sperm quality and fertility potential than aged roosters (p<0.05). Likewise, diet selenium supplements improved post-thaw sperm quality and fertility compared with the non-supplement group. Conclusion: Rooster's age does not influence the rooster sperm quality of fresh semen, while sperm cryotolerance and fertility were greater in young roosters than in aged roosters. However, sperm of aged roosters could be improved by dietary selenium supplementation.

• Animal Bioscience
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• v.36 no.12
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• pp.1796-1805
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• 2023

Objective: This study aims to identify heat shock protein70-2 (HSP70-2) and protamine-1 (PRM1) mRNA and protein in Madura bull sperm and demonstrate their relation as bull fertility biomarkers. Methods: The Madura bull fertility rates were grouped based on the percentage of first service conception rate (%FSCR) as high fertility (HF) (79.04%; n = 4), and low fertility (LF) (65.84%; n = 4). mRNA of HSP70-2 and PRM1 with peptidylprolyl isomerase A (PPIA) as a housekeeping gene were determined by quantitative real-time polymerase chain reaction, while enzyme-linked immunoassay was used to measure protein abundance. In the post-thawed semen samples, sperm motility, viability, acrosome integrity, and sperm DNA fragmentation index were analyzed. Data analysis was performed on the measured parameters of semen quality, relative mRNA expression, and protein abundance of HSP70-2 and PRM1, among the bulls with various fertility levels (HF and LF) in a one-way analysis of variance analysis. The Pearson correlation was used to analyze the relationship between semen quality, mRNA, proteins, and fertility rate. Results: Relative mRNA expression and protein abundance of HSP70-2 and PRM1 were detected and were found to be highly expressed in bulls with HF (p<0.05) and were associated with several parameters of semen quality. Conclusion: HSP70-2 and PRM1 mRNA and protein molecules have great potential to serve as molecular markers for determining bull fertility.

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.6
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• pp.705-709
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• 1996

A simplified dilutor for cock spermatozoa at ambient temperature was achieved by adjusting the 5% concentration of fructose in isotonic saline. Motility of cock spermatozoa was arrested completely for maximum 6 hrs without affection the survivability of spermatozoa by employing this sugar. To study the effect of high concentration of fructose on fertility, sperm were inseminated with or without fructose at different hrs. Fructose from semen samples was removed by centrifugation. High fertility obtained in the hens inseminated with fructose free sperm (washed). In addition, washed sperm maintained the 85.00% fertility for 6 hrs in winter season ($17-21^{\circ}C$) and 82.67% fertility for 3 hrs in summer season ($31-35^{\circ}C$). Whereas control groups showed 47.33 and 25.33% fertility in winter and summer season respectively. No significant difference was found in percent motility and live counts between the control and washed experimental groups during winter season. However, these measures differed significantly in summer. Washing of cock spermatozoa more than once, high speed centrifugation and more duration for centrifugation proved harmful to fertility. It may be concluded that fructose (5%) can be used as a motility or metabolic inhibitor of spermatozoa for short-term storage of cock semen at ambient temperatures.

• Reproductive and Developmental Biology
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• v.38 no.1
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• pp.53-62
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• 2014

The objective of this study was to investigate the relationship between fertility and protein pattern change using in vitro fertilization, analysis of sperm characteristics and two-dimensional gel electrophoresis in different pig types. In results, the viability and mitochondria integrity of sperm were higher significantly (p<0.05) but the portions of acrosome reaction was lower significantly (p<0.05) in Duroc and $F_1$ (potbellied ${\times}$ PWG miniature pig) than PWG miniature. On in vitro fertilization to investigate fertility, the fertility of $F_1$ semen war higher significantly (p<0.05) than in Duroc and PWG miniature pig. On the other hand, protein patterns showed similar function among the different boar semen. Especially, the heat shock 70 kDa 1-like and G patch domain-containing protein 4 were significantly (p<0.05) higher expressed in $F_1$ than in Duroc and PWG miniature pig. The proteins associated with mitochondria in Duroc were significantly (p<0.05) higher expressed than in $F_1$ and PWG miniature pig. The developmental rates to blastocyst stage of oocytes fertilized with sperm of $F_1$ pig were significantly (p<0.05) higher than in PWG miniature pig. However, phosphoglycerate kinase 2 and zinc finger protein 431 were significantly (p<0.05) higher expressed in PWG miniature pig than in $F_1$ and Duroc pigs. In conclusion, the results of the present study indicate that different proteins were expressed in different pig types, and were associated with a sperm functions and embryo development.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.3
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• pp.101-105
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• 2015

Objective: Several publications have established a relationship between sperm DNA damage and male factor infertility, based on data from America, Europe, and Asia. This study aimed to compare the extent of sperm DNA damage in sperm samples from Nigerian men with unexplained infertility and in sperm samples from a fertile group composed of sperm donors who had successfully impregnated a female partner naturally or through assisted conception. Methods: A total of 404 men underwent male fertility evaluation at Androcare Laboratories and Cryobank participated in this study. Semen analysis and a sperm chromatin structure assay (SCSA) were performed on all subjects. Results: The men in the unexplained infertility group were slightly older than the men in the fertile sperm group ($36{\pm}10$ years vs. $32{\pm}6$ years, p=0.051). No significant difference was observed between the two groups in semen analysis parameters ($p{\geq}0.05$). Men in the unexplained infertility group with normal semen parameters had a significantly higher DNA fragmentation index (DFI) than men in the fertile sperm group ($27.5%{\pm}7.0%$ vs. $14.1%{\pm}5.3%$, p<0.05). In the unexplained infertility group, 63% of the men had a DFI greater than 20%, compared to 4% in the fertile sperm group. In the unexplained infertility group, 15.2% of the subjects had a DFI greater than 30%, compared to 1% in the fertile sperm group. Conclusion: Our study showed that the SCSA may be a more reliable predictor of fertility potential than traditional semen analysis in cases of unexplained infertility.