• Journal of Embryo Transfer
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• v.15 no.3
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• pp.237-245
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• 2000

This study was carried out to elucidate whether sperm contain a factor inducing second polar body extrusion and to search for an effective collection method of the sperm factor Thus, sperm extract, dialyzed sperm-extract or liquid chromatographic fractions of sperm extract was microinjected into ovulated oocytes. And the microinjected oocytes were incubated for 24 hours to investigate about the extrusion of second polar body. The results obtained were as follows; 1. Sperm extract significantly increased the second polar body extrusion. 2. Sperm extract showed five major fractions at retention volumes (RVs) 1.25, 1.37, 1.84, 2.10 and 2.67ml after separation with Superose 12 column. These sperm extract fractions did not significantly increase the second polar body extrusion. 3. Dialyzed sperm-extract significantly increased the second polar body extrusion 4. Dialyzed sperm-extract showed three maior fractions at RVs 1.88, 2.14 and 2.77ml after separation with Superose 12 column. Of these fractions, the fraction RV2.14 significantly increased the second polar body extrusion. In conclusion, sperm extract contained a factor inducing the second polar body extrusion and the factor was contained largely in fraction RV2.14 after dialysis and liquid chromatographic fractionation of sperm extract.

• Clinical and Experimental Reproductive Medicine
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• v.49 no.2
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• pp.110-116
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• 2022

Objective: Sperm vitrification leads to the production of reactive oxygen species (ROS) that can damage the functional parameters of sperm. The present study aimed to investigate the antioxidant effect of Nigella sativa extract on motility, plasma membrane function, mitochondrial membrane potential (MMP), DNA damage, and intracellular ROS production. Methods: A total of 20 sperm samples were used. Samples were divided into six experimental groups, including groups with aqueous extract from N. sativa seeds at concentrations of 1% to 6%, a cryopreserved control group, and a fresh control group. Results: Statistical analysis showed significantly higher total sperm motility at concentrations of 3% to 6% than in the vitrified semen control group. Additionally, progressive motility and all motion characteristics at all concentrations were significantly higher than in the vitrified semen control group. The presence of N. sativa seed extract also improved the quality of the sperm parameters assayed in all experimental groups (1%-6%; intracellular ROS production, DNA damage, MMP, and sperm membrane function) compared to the control group. Conclusion: Higher concentrations of N. sativa led to improvements in all sperm parameters and sperm quality. These findings indicate that N. sativa seed extract is effective for improving the quality of sperm after vitrification.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.2
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• pp.60-66
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• 2019

Objective: To compare black rice (Oryza sativa L) extract with three different staining methods for human sperm head assessment. Methods: Semen samples were collected from 34 volunteers. Four smears of each ejaculate were prepared for staining using the rapid Papanicolaou (PAP) stain, SpermBlue, DipQuick, and black rice extract. The percentage of defective sperm heads (mean${\pm}$standard deviation) was compared. Results: Black glutinous rice extract, a natural dye, was used instead of hematoxylin to stain the nuclei of the sperm heads. The percentage of defective sperm heads showed a significant difference between black rice extract and DipQuick (p= 0.000). In contrast, black rice extract and rapid PAP showed no statistically significant difference (p= 0.974). A strong correlation (r = 0.761) was found between the findings obtained using rapid PAP and black rice extract. In contrast, a weak correlation (r = 0.248) was obtained between DipQuick and black rice extract for the percentage of defective sperm heads. Conclusion: The results showed good agreement and a strong correlation between the rapid PAP and black rice extract stains. The advantages of black rice extract as a novel substitute for hematoxylin for nuclear staining include ease of preparation, local availability, and favorable nuclear staining properties. Further studies could also focus on comparing staining techniques in clinical samples.

• Reproductive and Developmental Biology
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• v.37 no.1
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• pp.9-16
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• 2013

Artificial insemination (AI) has been performed widely in swine industry using fresh liquid sperm instead of frozen type of sperm. However fresh sperm are not able to preserve more than three days with optimal motility and other sperm parameters for the successful fertilization, since in vitro stored sperm has an oxidative stress that resulted increase of abnormality and acrosome reation. To overcome these major problems, novel preservative formulation is needed to neutralize the oxidative stress and to provide suitable physiological environment for sperm in in vitro. In this study, naturally derived substances such as Poncirus trifoliate (Trifoliate orange), Garcinia mangostana (Mangosteen), pig placenta and testis extracts were tested as sperm preservative agents. Placenta extracts (PE), trifoliate orange extracts (TOE), testes extracts (TE) and mangosteen extracts (ME) were applied to analyze specific parameters for sperm motion characteristics individually and combinatorial. Each individual extract treatment can accelerate the sperm motility but noticeably TOE, TE and ME treatments exhibited the considerable and significant preservation of sperm motility. PE, TE and ME showed a significant (p<0.05) increase in ALH after one week. Further we evaluated the five different combinations of these extracts on sperm motility and its motion characteristics. Surprisingly even after one week ME, TOE and TE combination significantly preserved the sperm motility about 75%. It is noteworthy that unlike individual extract treatment, combination of ME, TOE and TE simultaneously protect the sperm motility and its motion characteristics. Taken together these data conclude that addition of ME, TOE and TE can be effective for preservation of pig sperm.

• The Journal of Korean Medicine
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• v.27 no.3 s.67
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• pp.63-76
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• 2006

Objective: These studies were undertaken to evaluate the effects of the different administration duration of Morindae Radix extract solution on spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testes and the activities of sperm hyaluronidase. Materials and Method: We used 8-week-old ICR mice and administered 0.3mg/g extract solution of Morindae Radix once a day for 30, 60, 90 and 120 days. The control group was administered normal saline in the same way and duration. We examined the number of total, motile and normal sperm from the cauda epididymis. We also compared the testicular tissue, especially seminiferous tubules, between the control and treated groups by histochemical methods. Finally, we observed the difference of sperm hyaluronidase activities between the control and treated groups. Results: Significant administration duration-dependent differences were observed in the concentration of total sperm, motility and normality of spermatozoa of the Morindae Radix extract solution administered groups compared to the control group. In the histological analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the Morindae Radix extract solution administered groups compared to the control group. Also, the activity of hyaluronidase was significantly increased in the Morindae Radix extract solution administered groups compared to the control group. Conclusions: This study shows that the beneficial effect of Morindae Radix extract solution on the concentration, motility and morphology of sperm, the testicular tissues and the activities of sperm hyaluronidase increased the greater the duration the mice were administered it. We suggest that Morindae Radix may be useful for the treatment of male sexual dysfunction and infertility.

• Biomolecules & Therapeutics
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• v.21 no.2
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• pp.153-160
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• 2013

We screened the pharmacological effects of a 50% ethanol extract of Yacon tubers and leaves on spermatogenesis in rats. As a result, we found that Yacon tuber extracts increased sperm number and serum testosterone level in rats. It has been reported that the crude extract of Yacon tubers and leaves contain phenolic acids, such as, chlorogenic acid, ferulic acid and caffeic acid by HPLC/MS analysis. We were interested in the contributions made by phenolic acid, particularly chlorogenic acid of Yacon tuber extract to the spermatogenic activity. After administering Yacon tuber extract or chlorogenic acid to rats for 5 weeks, numbers of sperm in epididymis were increased by 34% and 20%, respectively. We also administered ferulic acid, which has been reported to be a metabolite of chlorogenic acid and a constituent of Yacon tuber extract to investigate its spermatogenic activity in rats. Yacon tuber extract and ferulic acid increased sperm numbers by 43% and 37%, respectively. And, Yacon tuber extract, and chlorogenic acid showed significantly inhibition effect of testoeterone degradation in rat liver homogenate. We considered that the spermatogenic effect of Yacon tuber extract might be related to phenolic compounds and their inhibitory effect of testosterone degradation. Yacon showed the possibility as ameliorable agents of infertility by sperm deficiency and late onset hypogonadism syndrome with low level of testosterone.

• The Journal of Korean Obstetrics and Gynecology
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• v.21 no.1
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• pp.16-27
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• 2008

Purpose: This study was conducted to investigate effects of Allii tuberosi Semen, Ginseng Radix Alba and Morindae officinalis Radix extract on reproductive capacities in mice. Methods: The 8-week-old ICR rats are used in the study, and we administered the extract solution of Allii tuberosi Semen, Ginseng Radix Alba and Morindae officinalis Radix concentration of 1, 10, 100mg/0.3ml to each rats and water to another rats(control group) once a day for each 30, 60, 90 and 120 days. we evaluated the total sperm count and morphology. Results: For 30 days, the total sperm count and normal sperm most increased in the group administered by 100mg/0.3ml Allii tuberosi Semen. For 60 days, the total sperm count and normal sperm most increased in the group administered by 100mg/0.3ml Morindae officinalis Radix solutions. For 90 days, the total sperm count most increased in the group asministered by 10mg/0.3ml Morindae officinalis Radix and 100mg/0.3ml Allii tuberosi Semen solutions. For 120 days, the total sperm count and normal sperm most increased in the group administered by l0mg/0.3ml Ginseng Radix Alba. Conclusion: This study shows that there were dose and duration effects of Allii tuberosi Semen, Ginseng Radix Alba and Morindae officinalis Radix extract solution on the total sperm count and morphology.

• Journal of Embryo Transfer
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• v.12 no.2
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• pp.219-226
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• 1997

Follicular fluid influxed into the oviduct during ovulation may affect movement of sperm for fertilization Thus, in this study, the effect of follicular fluid, obtained from follicles of l0mm in diameter, on number and quality of sperm recovered by swim-up separation was investigated and sperm-movement stimulating components extracted from follicular fluid with methanol and isooctane were separated by gel filtration with Sepadex G-1O, G-25 and G-1OO gels, and were isolated by electrophoresis with SDS-PAGE mini gel. The results obtained were as follows; 1. Diluted follicular fluid stimulated sperm movement. 2. Sperm-movement stimulating factors were in methanol extract. 3. Sperm-movement stimulating effect of methanol extract appeared in fraction I among fractions recovered after gel filtration. And the fraction I contained proteins indicating 4 major bands as about 47, 43, 25 and 14 kilodaldons and 5 minor bands as about 67, 58, 23, 22 and 21 kilodaldons. 4. The fraction I recovered from G-100 gel showed significantly low percentage of motile sperm and had no protein indicating the band of 67 kilodaldons among the minor bands.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.3
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• pp.17-31
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• 2005

Purpose : These studies were undertaken to evaluate the effects of the administration of different concentrated Morindae officinalis Radix extract solution on the spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testis and the activities of sperm hyaluronidase, testicular peroxidase and testicular catalase. Methods : We used the 2-month-old mice and administered the extract solution of Morindae officinalis Radix in the different concentration once a day for 60 days. The control group was administered the normal saline in the same way and duration. We examined the number of total, motile and normal sperm from the cauda epididymis, the activities of sperm hyaluronidase, testicular peroxidase and testicular catalase. Also we observed changes of isolated testis before and after administration of Morindae officinalis Radix extract solutions the mice. And we compared to the testicular tissue especially seminiferous tubules between control and treated group by histochemical methods. Results : The significant dose-dependent differences were observed in the concentration of total sperm, the motility and normality of spermatozoa of Morindae officinalis Radix extract solution administered groups compared to the control group, respectively. In the histological analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the Morindae officinalis Radix extract solution administered groups compared to the control group, respectively. Also, the activity of hyaluronidase was significantly increased in the Morindae officinalis Radix extract solution administered groups compared to the control group. In the antioxidant activity analysis, the activity of testicular peroxidase was significantly increased in the Morindae officinalis Radix extract solution administered groups compared to the control group, respectively. Conclusion : This study shows that Morindae officinalis Radix has the beneficial effect on the concentration, morphology and motility of sperm, the activities of sperm hyaluronidase and testicular peroxidase. We can suggest that Morindae officinalis Radix extract solution be useful for the treatment of male sexual dysfunctions and infertility.

• Animal Bioscience
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• v.37 no.7
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• pp.1177-1184
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• 2024

Objective: Semen cryopreservation is an effective method of preserving genetic material, particularly in native chicken breeds facing a substantial decline. In this study, we evaluated the quality of frozen/thawed rooster semen treated with different concentrations of oral administrations of black ginger (Kaempferia parviflora: KP) extract and determined its fertility. Methods: Thirty-two Thai native roosters (Pradu Hang Dum, 42 weeks old) were used in this study. The treatments were classified into four groups according to the concentration of KP extract administered to the roosters: 0, 100, 150, and 200 mg/kg body weight. The quality of fresh semen was analyzed before cryopreservation. Post-thaw sperm quality and fertility potential were determined. Also, lipid peroxidation was determined. Results: The results showed that sperm concentration and movement increased in roosters treated with 200 mg/kg of KP extract (p<0.05). The malondialdehyde (MDA) in the roosters receiving 200 mg/kg KP extract was lower than that in the other but had an insignificant difference within the KP treatment groups (p>0.05). The highest MDA levels were observed in the control group (p<0.05). The percentage of motile sperm (total motility and progressive motility) after semen thawing was higher in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (p<0.05). MDA levels decreased significantly in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (p<0.05). Fertility and hatchability were greater in the KP150 and KP200 groups than in the KP100 and control groups (p<0.05). Conclusion: The optimal amount of KP extract influencing initial sperm quality was determined to be 200 mg/kg. However, 150 mg/kg was the optimal low dosage of KP extract administration that maintained sperm quality and fertility following semen cryopreservation.