• Journal of Veterinary Clinics
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• v.21 no.4
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• pp.329-335
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• 2004

Dog spermatozoa were recovered from the caudae epididymides of 23 domestic dogs which were 11 pure breed and 12 mix-breed dogs ranging in age from 0.6 to 3 years. The experimental designs were as follows: 1) the effect of chilling to 0. 10 or 37$^{\circ}C$. 2) the kinetics of chilling injury at 0 or 4$^{\circ}C$, and 3) the effect of sugars at $0^{\circ}C$. Viable spermatozoa were recovered by percoll gradient separation and adjusted to 5${\times}$10$^{7}$ spermatozoa/ml. In experiment 1, spermatozoa were diluted with 0.33 M glucose supplemented with 3% BSA (G-BSA) at 1:2 dilution. Spermatozoa were loaded into straws and exposed at 0, 10 or 37$^{\circ}C$ for 30 min. In experiment 2, spermatozoa were prepared as the experiment 1 and exposed for 0.5, 5, 15, or 30 min at 0 or 4$^{\circ}C$. In experiment 3, spermatozoa were diluted with different sugars (0.33 M galactose, glucose, fructose, mannitol, lactose, sucrose, raffinose) and cooled to $0^{\circ}C$ for 30 min. Sperm membrane integrity, motility and acrosome integrity were assayed after rewarming at 37$^{\circ}C$ for 5 min. Sperm motility and membrane integrity abruptly decreased with decreasing temperature but acrosome integrity gradually decreased (P<0.05). Sperm motility was more sensitive to cold shock than membrane integrity and acrosome integrity. Spermatozoa cooled to $0^{\circ}C$ were more damaged than those at 4$^{\circ}C$. Sperm motility was not different among exposed times at both. 0 and 4$^{\circ}C$. However, membrane integrity of spermatozoa exposed for 30 min at both 0 and 4$^{\circ}C$ was significantly lower (P<0.05). Spermatozoa diluted in 0.33 M fructose or galactose showed lower motility and higher morphological abnormality with coiled tail at $0^{\circ}C$. These sperm characteristics were strongly related. These results indicate that dog epididymal spermatozoa are relatively sensitive to rapid cooling and higher morphological abnormality at $0^{\circ}C$ was shown in spermatozoa diluted in fructose and galactose.

• Asian-Australasian Journal of Animal Sciences
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• v.5 no.3
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• pp.449-454
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• 1992

Eight healthy Holstein bulls, 4-6 years old were used to study the effect of season of the year on the incidence of the morphologically abnormal spermatozoa. Semen was collected twice a week by A. V. over one-year period. The percentage of total abnormal spermatozoa was $14.1{\pm}0.5$. Ejaculates collected during hot summer season had significantly higher incidence of abnormal spermatozoa than those collected during winter time. Warm spring had moderate semen abnormality. In addition to its effect on the total number of morphologically abnormal spermatozoa, season affected significantly the primary as well as the secondary types of abnormalities. The differences between incidence of primary and secondary types of abnormalities were not significant for all seasons and seasons pooled together. The ratio between the total forms of abnormality in the head, mid-piece and tail of spermatozoa was as 1:1.5:1. Head and mid-piece had defected more during summer compared with both winter and spring. There was no variation in tail abnormalities due to season. The significant effect of season on head was observed by large, pyriform, free and detached heads, while that on mid-piece was by swollen, coiled mid-pieces and protoplasmic droplets.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.1
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• pp.80-84
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• 2021

Objective: The aim of the study was to investigate pregnancy, obstetric, and neonatal outcomes in women with small (<4 cm) unilateral endometriomas. Methods: This retrospective study included 177 patients: 91 patients with small endometriomas and 86 controls with unexplained or tubal factor infertility who were treated at the Süleymaniye Gynecology and Maternity Training and Research Hospital Infertility Unit between January 2010 and July 2015. The groups were matched with regards to demographic characteristics such as age, body mass index, and infertility duration. All of the women in this study conceived via intracytoplasmic sperm injection. We compared pregnancy, obstetric, and neonatal outcomes between these groups. Results: Women with endometriomas had a higher biochemical pregnancy rate, but lower clinical pregnancy and live birth rates than women with unexplained and tubal factor infertility (p<0.05 for all). However no significant differences were found in terms of obstetric and neonatal complications between the two groups (p>0.05 for all). Conclusion: In this study, we found that women with endometriomas less than 4 cm were more prone to early pregnancy complications. We also showed that this group did not have any increased risks of late pregnancy, obstetric, and neonatal complications.

• Proceedings of the KOSOMBE Conference
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• v.1992 no.11
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• pp.15-17
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• 1992

정자 운동의 분석은 붙임의 원인 규명과 진단에 중요한 정보를 제공한다. 본 논문에서는 연속된 정자 운동 영상을 이용해 정자 운동성의 주요 척도인 선형속도, 곡선속도, 선형성, 운동력등의 운동특성을 검출해 내는 알고리즘을 구현했다. 비디오 신호로 보내진 정자 영상을 컴퓨터에 의해 디지털화하고, 각 정자 머리의 위치를 계산해내었고, 이로부터 정자 등의 경로 및 주요 운동특성를 찾았다.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.81-87
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• 2007

The purpose of the present study was to determine the preventive effects of the two antioxidant vitamin E and catechin on DEHP-induced disturbance of spermatogenesis in male rats. Rats at 4 weeks of age were randomly allocated into five groups with 20 animals per group. The first group was not any administrated as control. The second group was administrated DEHP (2 g/kg) daily for 14 days. The third group was administrated vitamin E (500 IU/kg) following DEHP treatment by the same method (daily for 14 days). The fourth group was administrated catechin (200 mg/kg) following DEHP treatment by the same method. The fifth group was co-administrated vitamin E (500 IU/kg) and catechin (200 mg/kg) following DEHP treatment by the same method. In order to determine the preventive effects, we examined pathological changes of testis with apoptotic index, and characteristics of sperm with computer assisted sperm analysis (CASA). Vitamin E and catechin supplementation were significantly prevented the testicular atrophy, apoptosis of germ cells in the seminiferous tubules and abnormal rate of sperm. Moreover, sperm concentration, viability and motility was significantly recovered in groups of alone and along with vitamin E and catechin. The results suggest that preventive effects of alone and along administration of vitamin E and catechin on DEHP-induced testicular atrophy damages have been demonstrated.

• Korean Journal of Veterinary Service
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• v.40 no.3
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• pp.201-208
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• 2017

BVDV causes significant infections in ruminants, resulting in reproductive disorders, diarrhea, reduced milk production and enormous damage to farms. In particular, identification and culling of persistent infectious calf is an important task to eliminate infectious nidus in cattle households. However, studies on physiological characteristics of PI bull are still insufficient to understand reproductive effects of BVDV. In this study, one PI bull was confirmed in herd and complete blood analysis was performed. The lymphocyte count of PI at age 4 was below the normal range and the number of WBCs was also in the lower level of normal range in blood. The sperm number produced by PI male becomes lower and the viability of fresh sperm comes to poor with ages (P<0.05). The sperm abnormality was also increased, especially in nuclear vacuoles of head and droplets of midpeace (P<0.05). The PI male becomes infertile due to poor semen quality at age 4. With these results, we concluded that BVDV in PI bull cause decreased sperm cell and abnormality in semen so causes infertility. However, it appears that BVDV could not be transmitted by indirect contact of PI bull, because there was no evidence of BVDV infection in the herd, when regular vaccination program was applied.

• Reproductive and Developmental Biology
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• v.30 no.1
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• pp.59-64
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• 2006

This study was conducted to establish a convenient freezing method of boar semen. Boar semen was cooled until $5^{\circ}C$ for 3 hrs using cell freezer and loaded into straws. Semen straws were frozen in different steps in strofoam box filled with $LN_2$. Highest sperm viability (54.0%) was obtained by 1-step freezing(holding at 10 cm height from the surface of $LN_2$ for 10 min). Sperm viability increased by holding at $-102^{\circ}C$ for 10min (74.0%, P<0.05). In thawing regime, sperm viability was significantly higher in $37^{\circ}C$ group than in $52^{\circ}C$ group. The sperm characteristics did not differ between 1-step and 3-step. After IVF using frozen-thawed boar semen, developmental rate of embryos to the morula+blastocyst stage was in 1-step freezing group than that of 3-step freezing group (27.5 vs 14.7%, P<0.05). The result shows that the 1-step freezing with holding at $-102^{\circ}C$ for 10min before plunging into $LN_2$ is a convenient and easy freezing method for boar semen.

• Korean Journal of Environment and Ecology
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• v.35 no.5
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• pp.503-524
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• 2021

A largemouth bass (Micropterus salmoides) is an ecosystem disturbance fish species at the highest rank in the aquatic ecosystem, causing a serious imbalance in freshwater ecosystems. Although various attempts have been made to eradicate and control largemouth bass, no effective measures were found. Therefore, it is necessary to find an approach to maximize the effective population reduction based on the unique characteristics of largemouth bass. This study used the transcriptome analysis to derive 182,887 unigene contigs and select 12 types of final target sequences for applying the CRISPR/Cas9 system in the genes of IZUMO1 and Zona pellucida sperm-binding protein, which are proteins involved in sperm-egg recognition. After synthesizing 12 types of sgRNA capable of recognizing each target sequence, 12 types of Cas9-sgRNA ribonucleoprotein (RNP) complexes to be used in subsequent studies were prepared. This study searched the protein-coding gene of sperm-egg through the Next Generation Sequencing (NGS) and edited genes through the CRISPR/Cas9 system to induce infertile individuals that produced reproductive cells but could not form fertilized eggs. Through such a series of processes, it successfully established a composition development process for largemouth bass. It is judged that this study contributed to securing the valuable basic data for follow-up studies to verify its effect for the management of ecological disturbances without affecting the habitat of other endemic species in the same water system with the largemouth bass.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.19 no.9
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• pp.116-123
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• 2018

The sperm quality is determined by the kinetic characteristics and acrosome integrity of the sperm. In the previous studies, analysis of semen quality had large errors because those experiments by using microscope had been conducted by people. In recent years, the molecular biological methods have been newly developed to complement the previous techniques. The ZAR1 gene is known to be a gene that affects early embryonic development in vertebrates, but there is no study of the association with semen. In this study, we analyzed the association between the kinetic characteristics and ZAR1 single nucleotide polymorphism (SNP) genotype. To detect the SNPs, we performed sequencing using genomic DNA from the whole bloods of Duroc pigs. We identified an SNP in the ZAR1 gene g.2540T>C. ZAR1 SNP genotypeing in 105 pigs revealed that the major and minor alleles were T and C, respectively. After we analyzed the association between the kinetic characteristics of sperm and the ZAR1 SNP genotype, we found a significant association in MOT (p<0.01), VSL (p<0.05) of the kinetic characteristics in the Duroc boars. It was confirmed that the boars with T allele were lower in MOT and VSL than C allele. Therefore, pigs with C allele are judged to be better at the MOT and VSL of semen. Based on these results, ZAR1 SNP genotyping may be a useful molecular biomarker to improve semen quality by applying molecular breeding technology.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.10
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• pp.1276-1281
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• 2010

The present study was conducted to investigate the effects of butylated hydroxytoluene (BHT) supplementation on diluted, cooled and frozen-thawed ram spermatozoa. After primary evaluation of collected ejaculates, only semen samples with motility of more than 70% and sperm concentration higher than $3{\times}10^3$ sperm/ml were used for cryopreservation. The selected semen samples were then pooled and diluted 1:4 with Tris Citrate Fructose Yolk (TCFY) extender supplemented with different concentrations of BHT (0.5, 10, 2.0 and 3.0 mM). As the control, semen was diluted and frozen in the diluent without BHT. Motility, progressive motility, viability, membranes and acrosome integrity were evaluated after dilution (part 1), cooling (part 2) and freezing and thawing (part 3). The results of the first part of the experiment showed that there were no significant difference between treatments in the motility, progressive motility, viability, membranes and acrosome integrity of spermatozoa, but the results with 2.0 mM BHT were slightly better than obtained with other levels of BHT and control extender. Significantly better results (p<0.05) were observed in the second part of the experiment for cooled spermatozoa characteristics, when extender was supplemented with 2.0 and 3.0 mM BHT. Furthermore, the results obtained in the third part of the experiment indicated that, after freezing and thawing, all evaluated semen characteristics were improved significantly (p<0.05) by increasing BHT levels, with the best results obtained for extender containing 2 mM BHT. Comparison of these results with those of control diluent, the effects of supplementation were significantly (p<0.01) better. However, the higher concentration of BHT (3.0 mM) reduced the motility, acrosomal integrity, viability and hypo-osmotic swelling response of spermatozoa compared to extender containing 2.0 mM BHT. In conclusion, the results obtained in this study showed that the semen quality of rams was improved when BHT was added to extender used before the freezing process.