• Journal of Aquaculture
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• v.19 no.1
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• pp.52-56
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• 2006

This study was conducted to investigate the effects of three different concentrations (6%, 8% and 10% final volume) of dimethyl sulfoxide (DMSO) on cryopreserved sperm of grass carp (Ctenopharyngodon idellus). Grass carp sperm was suspended in Kurokura extender #2 and equilibrated at $4^{\circ}C$ for 10 min. French straws (0.25 ml) of sperm were frozen from $4^{\circ}C\;to\;-4^{\circ}C$ at a rate of $4^{\circ}C\;min^{-1}$ and then ken $-4^{\circ}C\;to\;-80^{\circ}C$ at a rate of $11^{\circ}C\;min^{-1}$. The straws were kept at $-80^{\circ}C$ for 10 min and finally stored in liquid nitrogen $(-196^{\circ}C)$. The cryopreserved sperm was thawed in a water bath at $40^{\circ}C$ for 30 sec and fertilization, hatching rate and larval malformation were compared with fresh sperm (control). The fertilization rate of post-thawed sperm was comparable (from 88.21% to 94.30%) to that of fresh sperm. However, hatching rate of all frozen sperm were significantly lower (P<0.05) than that of control. Additionally, the larval abnormality rate of frozen sperm was significantly higher than that of fresh sperm. The results indicate that DMSO could affect the quality of cryopreserved sperm of grass carp, and a freezing program and a proper extender composition should be further studied.

• Clinical and Experimental Reproductive Medicine
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• v.16 no.2
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• pp.173-177
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• 1989

Defective or inadequate semen quality, usually presenting as low sperm count or poor sperm motility , is recognizable by semen analysis. However, the ability of spermatozoa to fertilize an ovum is not determined used in various experiments. In this study, hamster oocyte sperm penetration assay was used to determine the fertilizing capacity of sperms in 20 subjects which divided into two groups, group A with 10 normal fertile men, and group B with 10 infertile men. The % penetration in group A and group B were 61% and 35% respectively, which showed statistically not significant but fertilization index was significantly different between group A(FI=2.24) and group B(FI=O.05). Additionally it seemed that the percentage of sperm penetraton was influenced more by the motility of spermatozoa than by the number.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.2
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• pp.190-197
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• 2008

This study was designed to analyze boar sperm to compare motility, acrosome morphology, viability and ATP by various preservation methods between Duroc boar A with cold shock resistance sperm and Duroc boar B with cold shock sensitivity sperm. Semen volume, sperm concentration, motility and normal acrosome between Duroc boar A and B did not show any differences within 2 h after collection. There were no differences in sperm motility and normal acrosome between boar A and B at 1 day of preservation at $17^{\circ}C$ and $4^{\circ}C$, respectively. However, sperm motility and normal acrosome from 2 day of preservation at $17^{\circ}C$ and $4^{\circ}C$, respectively, were higher for boar A than boar B. The frozen-thawed sperm motility and normal acrosome were higher for boar A than boar B. The sperm viability and ATP concentration according to storage period of liquid semen at $17^{\circ}C$ and $4^{\circ}C$ were higher for boar A than boar B. Also, the sperm viability and ATP concentration of frozen-thawed semen were higher for boar A than boar B. In conclusion, we found out that the original quality of boar semen with cold shock resistance sperm played an important role.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.1
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• pp.10-17
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• 2011

Objective: To explore potential relationships between sperm DNA integrity and both semen parameters and clinical outcomes. Methods: Semen analysis of 498 samples was performed according to the 2010 criteria of the World Health Organization. The sperm DNA fragmentation Index (DFI) of the semen samples was assessed using a neutral comet assay. Results: Sperm DFI showed a significant correlation with semen parameters, including the patient's age, sperm viability, motility, morphology, and number of leukocytes (p<0.05). The sperm DFI values for asthenozoospermic (15.2%), oligoteratozoospermic (18.3%), asthenoteratozoospermic (17.5%), and oligoasthenoteratozoospermic semen samples (21.3%) were significantly higher than that observed in normozoospermic semen samples (10.5%, p<0.05). A sperm DFI value of 14% was used as a threshold of sperm DFI in assessing whether DNA was highly damaged. In 114 IVF-ET cycles, the fertilization rate of the sperm DFI <14% group (70 cycles, 61.7%) was significantly higher than that observed for the ${\geq}14%$ group (44 cycles, 55.3%), but there was no difference in the other clinical outcomes between the two groups. In the ${\geq}14%$ group, the pregnancy rates of the ICSI cycles (40.0%) and half-ICSI (44.0%) were higher than conventional IVF cycles (30.7%), but the difference was not statistically significant. Conclusion: Along with the conventional semen analysis, the sperm DFI assessed using the comet assay was shown to improve the quality of the semen evaluation. To evaluate the precise effect of ICSI on pregnancy rates in the patients who demonstrate high sperm DFI values, further study is necessary.

• Animal Bioscience
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• v.35 no.9
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• pp.1351-1359
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• 2022

Objective: The objective of this study was to analyse the differentially abundant proteins caused by freeze-thawing of ram sperm and explore candidate proteins of interest for their ability to improve ram sperm cryopreservation outcomes in vitro. Methods: Sperm were from three mature Dorper. Fresh and frozen sperm proteins were extracted, and the differentially abundant proteins were analysed by mass spectrometry. Among these proteins, lactoferrin (LTF) was selected to be added before cryopreservation. Next, sperm samples were diluted in Tris extender, with the addition of 0, 10, 100, 500, and 1,000 ㎍/mL of LTF. After thawing, sperm quality was evaluated by motility, plasma membrane integrity, mitochondrial activity and reactive oxygen species (ROS). Results: Cryopreservation significantly altered the abundance of 40 proteins; the abundance of 16 proteins was increased, while that of 24 proteins was decreased. Next, LTF was added to Tris extender applied to ram sperm. The results showed that sperm motility and plasma membrane integrity were significantly improved (p<0.05) by supplementation with 10 ㎍/mL LTF compared to those in the control group. There was no significant difference in mitochondrial activity between the 0 ㎍/mL group and other groups (p>0.05). Supplementation of the cryoprotective extender with 10 ㎍/mL LTF led to decreased ROS levels compared with those in the control and other groups (p<0.05). Conclusion: The LTF is an important protein during cryopreservation, and the addition of 10 ㎍/mL LTF to a cryoprotective extender can significantly improve the function of frozen ram sperm.

• Animal Bioscience
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• v.36 no.12
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• pp.1821-1830
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• 2023

Objective: This study investigated the effect of adding seminal plasma to frozen-thawed semen on the quality of sperm and pregnancy following insemination in dromedary camels. Methods: In experiment 1, the frozen-thawed semen from 9 collections (3 bulls) was further diluted with either the base extender or homologous seminal plasma (HSP). In the second experiment, a pooled sample of frozen-thawed semen was diluted with either seminal plasma from another three bulls. Live percentage, total and progressive motility, functional and acrosome integrity, and sperm kinematics were evaluated at 15, 60, and 120 minutes post-thawing and compared to the non-treated control. In experiment 3, frozen semen was used to inseminate camels in the following experimental groups: 1-Single insemination with double dose undiluted frozen semen (n = 9); 2-Re-insemination in 6 hours with undiluted semen (n = 13); 3-Single insemination with HSP treated sperm (n = 14). Results: Frozen-thawed sperm diluted in HSP or the non-homologous seminal plasma from Bull C indicated an improvement in all parameters after 1 hour post-thawing incubation (p<0.05). The proportion of total and progressively motile sperm did not drop significantly at 60 minutes post-thawing when diluted with the seminal plasma of Bull C (p>0.05). Double insemination with nontreated sperm and single insemination with HSP-treated sperm resulted in similar pregnancy rates (15.3% vs 21.4%, p>0.05). None of the camels conceived with double-dose single insemination of nontreated sperm. Conclusion: Seminal plasma improves sperm longevity and motility after thawing in dromedary camel with a significant between-bull variation in effect. Low post-thaw sperm longevity might be the cause behind the low pregnancy rates in frozen semen insemination of dromedary camels.

• Advances in nano research
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• v.11 no.6
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• pp.649-655
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• 2021

Nanotechnology is widely considered a major technology of the twenty-first century. Nanoparticles (NPs) has been shown to pass through reproductively significant biological barriers such as the blood-testicle and placental barriers. Thus, the purpose of this study was to determine the effect of silver Nanoparticles (Ag-NPs) on sperm-egg interaction and spermatozoa quality parameters in quail spermatozoa. Semen was suspended in Ringer solution containing Ag-NPs levels at 5.5 × 106 sperm/ml (0, 0.01, 0.1, 1 and 10 ppm). The results indicated that when sperm were counted at 0.1 ppm, the number of holes formed on the inner perivitelline layer was significantly increased compared to the control. The 10 ppm group had a significant reduction in sperm viability. At 0.1 and 1 ppm, the membrane integrity was significantly decreased (P < 0.05). All treatments (except 0.01 ppm Ag-NPs) had a significant (P < 0.05) effect on the percentage of spermatozoa with an intact acrosome when compared to the control group. At 0.1, 1, and 10 ppm Ag-NPs, morphological defects in the acrosome were observed. As a result, Ag-NPs is likely capable of destroying the acrosome membrane. This research indicates that Ag-NPs may be cytotoxic to spermatozoa by impairing sperm functionality and increasing sperm mortality.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.4
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• pp.486-494
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• 2006

In order to protect the spermatozoa against cold shock, hen egg yolk is widely used as a cryoprotective agent in semen freezing extenders for domestic animals. The protective action of yolk is largely presumed to be due to low density lipoproteins (LDL). The effects of LDL on sperm quality of bull and northern pike (Esox lucius) after freezing-thawing have been reported, but no study has been made to evaluate the effect of LDL on boar sperm motility and other characteristics. The experiment was carried out to investigate the effect of LDL on the freezing of boar sperm in 0.25 ml straws. The aim was to evaluate the quality of boar spermatozoa cryopreserved in the presence of LDL. Motility of semen cryopreserved in LDL was analyzed and compared to semen cryopreserved with Tris-citric acid-glucose (TCG) and Tris-citric acid-fructose (TCF), two basic freezing extenders containing egg yolk. Similarly, acrosome and plasma membrane integrity were also evaluated and compared to semen cryopreserved with TCG and TCF. Analysis of sperm quality after freeze-thaw showed that the motility, acrosome and plasma membrane integrity were improved with LDL in the extender, as compared to the TCG and TCF. The highest post-thaw integrity of acrosome and plasma membrane and motility were obtained with 9% LDL (w/v). Consequently, the optimum LDL concentration in the extender was 9%. It is also suggested that the concentration of LDL addition is important for the effect on boar sperm protection during freezing and thawing. The percentage of motile spermatozoa was significantly higher after freezing in 9% LDL than in TCG and TCF 54.4% versus 30.4% and 30.1% (p<0.05), respectively. The integrity of acrosome and plasma membrane were also significantly higher at 70.3% and 50.5% respectively with semen frozen in 9% LDL extender compared to TCG at 37.8% and 30.3% and TCF at 36.4% and 29.9%, respectively (p<0.05),. In conclusion, we propose that extender containing LDL extracted from hen egg yolk could be used as a cryoprotective media with a better efficiency than TCG and TCF. LDL improved boar semen quality, allowing better spermatozoa motility, acrosome and plasma membrane integrity after the freeze-thaw process. Furthermore, we found out that the extender with 9% LDL concentration significantly enhanced motility, acrosome and plasma membrane integrity of boar sperm after freezing and thawing.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.587-593
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• 2021

Infertility is a key issue affecting mood and behavior in men. Microorganisms are one of the primary etiological agents that may be associated with infertility. The objective of the present study was to identify bacterial causative agents from the semen of infertile subjects and determine the effect of bacterial infection on sperm quality, as well as determine the susceptibility of these bacteria to drugs. Forty semen samples from 30 infertile patients and 10 fertile individuals were collected. The pH, volume, motility, and concentration of semen were analyzed. The samples were processed and identified by biochemical testing using API identification kits. The antibiotic susceptibility pattern was determined using the disc diffusion method. Abnormal sperm quality was observed. The mean age of the individual and their sperm morphology, concentration, progressive motility, pH level, and pus cell content were 31.9 years, 2.7%, 10.4 million/ml, 27.3%, 8.3, and 5.7, respectively. Among the tested samples, oligoasthenozoospermia was found to show the highest occurrence, at 27/30 samples, followed by teratozoospermia, at 25/30 samples, and asthenozoospermia, at 22/30 samples. Of the tested infertile patients' sperm, 19, 6, and 5 isolates were identified as Escherichia coli, Klebsiella pneumonia, and Staphylococcus epidermidis, respectively. The results also revealed multi-drug resistance in the bacteria. Compared to that shown by the other tested antibiotics, amikacin showed higher activity against all isolated bacteria. However, the bacteria exhibited maximum resistance against gentamicin, cefotaxime, levofloxacin, and ampicillin. In conclusion, leukocytospermia and bacterial infections are possibly responsible for sperm abnormalities. Multi-drug resistant bacteria were detected. Gentamicin, cefotaxime, levofloxacin and ampicillin were shown the highest resistance, while amikacin was the most effective antimicrobial agent against the isolated bacteria.