• Journal of Embryo Transfer
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• v.16 no.2
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• pp.133-138
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• 2001

The aim of this study was to identify the method on extended canine semen exposed to freezing as assessed by motility, survival and acrosomal changes following different freezing ramp rates. Five ejaculates collected by digital manipulation twice weekly from three dogs (Shih-Tzu) were added Tris-Egg Yolk (TE) buffer and divided into 4 aliquots according to formulation of our laboratory. After cooling to 4$^{\circ}C$ by ramp rate of 0.6$^{\circ}C$/min, the samples frozen by ramp rates of 1.6$^{\circ}C$/min to -$25^{\circ}C$, 3$^{\circ}C$/min to -35$^{\circ}C$, 8.9$^{\circ}C$/min to -7$0^{\circ}C$ and 19$^{\circ}C$/min to -11$0^{\circ}C$, respectively, and then stored in L$N_2$for 2days. Each sample was evaluated on their motility, survivability and acrosome integrity at different thawing temperature. The ramp rate of 3$^{\circ}C$/min to -35$^{\circ}C$/h for freezing and thawing temperature of 37$^{\circ}C$ obtained the highest results to improve survivability, motile spermatozoa and intact acrosome appearance than other onditions. In conclusion, we may suggest freezing semen for canine artificial insemination is more efficient with freezing at a ramp rate of -3$^{\circ}C$/min to -35$^{\circ}C$ and thawing with a water bath adjusted to 37$^{\circ}C$.

• Journal of Embryo Transfer
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• v.28 no.4
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• pp.325-335
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• 2013

The study was set for one year to measure the effects of concentrate supplementation on reproductive performances and semen quality in indigenous rams. The study was conducted at the Department of Surgery and Obstetrics, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh during the period from May 2011 to April 2012. Forteen ram lambs (4~5 months) were randomly divided into two equal groups (n=7); supplemented vs control. The animals of control group were maintained on natural grazing. Along with natural grazing the supplemented group was on supplemented feeding. The concentrate supplementation (Wheat bran, Crushed maize, Soy bean meal, Fish meal, DCP powder, Vitamin mineral premix, Salt) was provided @ 300 g/head /day to the supplemented group. Body weight, scrotal circumference, BCS and libido index were measured weekly. Age, body weight and scrotal circumference at puberty were recorded. Semen was collected once in a weak using artificial vagina and chilled at $5^{\circ}C$ for 48h for evaluation. Concentrate supplementation did not influence (p>0.05) body condition score, age, weight, scrotal circumference at puberty and libido index. Final body weight (kg), growth rate (g/d), scrotal circumference (cm) and scrotal growth rate (mm/15d) were significantly (p<0.05) higher in supplemented group of rams compared to control. Volume, concentration, motility and membrane potentiality of spermatozoa were varied significantly (p<0.05) in supplemented and control groups. However, density, mass motility, viability and sperm with normal acrosome, midpiece and tail were not differed insignificantly (p>0.05) in different observation times. It was concluded that concentrate supplementation with free grazing improved weight and scrotal circumference gain and semen production with increased quality in indigenous ram.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.3
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• pp.112-118
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• 2019

Objective: Oxidative stress contributes to male infertility, and antioxidants have been recommended for treating idiopathic oligoasthenoteratozoospermia (OAT). There is, however, a lack of agreement on the type, dosing, and use of individual antioxidants or combinations thereof. The purpose of this study was to compare the effects of two doses of coenzyme Q10 (CoQ10) on semen parameters and antioxidant status in men with idiopathic OAT. Methods: In this prospective study, patients with idiopathic OAT received 200 mg/day (n = 35) or 400 mg/day (n = 30) of CoQ10 orally for 3 months. All patients underwent semen analysis according to the fifth editions of the World Health Organization criteria. Total antioxidant capacity (TAC), catalase (CAT) activity, and superoxide dismutase (SOD) activity were measured both before and after treatment. Results: Treatment with CoQ10 (200 mg/day or 400 mg/day) resulted in a significant increase in sperm concentration from baseline ($8.22{\pm}6.88$ to $12.53{\pm}8.11million/mL$, p= 0.019; $7.58{\pm}5.41$ to $12.33{\pm}6.1million/mL$, p= 0.002, respectively), progressive motility ($16.54%{\pm}9.26%$ to $22.58%{\pm}10.15%$, p=0.011; $14.22%{\pm}12.85%$ to $26.1%{\pm}14.52%$, p= 0.001, respectively), and total motility ($25.68%{\pm}6.41%$ to $29.96%{\pm}8.09%$, p= 0.016; $23.46%{\pm}12.59%$ to $34.82%{\pm}14.17%$, p= 0.001, respectively). CoQ10 therapy also increased TAC (p= 0.009, p= 0.001, respectively), SOD activity (p= 0.004, p= 0.001, respectively), and CAT activity (p= 0.039, p= 0.024, respectively). Furthermore, antioxidant measures correlated significantly with seminal fluid parameters (r = 0.36-0.76). Conclusion: CoQ10 supplementation improved semen parameters and antioxidant status in men with idiopathic OAT, with a greater improvement shown in men who took 400 mg/day than in those who took 200 mg/day.

• Clinical and Experimental Reproductive Medicine
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• v.49 no.1
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• pp.33-39
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• 2022

Objective: Male genital tract infections have been associated with infertility, and Escherichia coli has drawn increasing attention as an important bacterium in this context. This investigation aimed to characterize and compare the distributions of O-antigen serogroups of E. coli in the semen samples of fertile and infertile men. Methods: In this case-control study, semen samples were collected from 618 fertile and 1,535 infertile men. The E. coli-positive samples were evaluated in terms of concentration, morphology, viability, and motility parameters according to the World Health Organization 2010 guidelines. Finally, different serogroups of E. coli were identified by multiplex polymerase chain reaction targeting the O-antigen variations of the bacterium. Results: The prevalence of E. coli among fertile men was significantly higher than among infertile men (p<0.001). The sperm morphology, viability, and motility in the E. coli-positive fertile group were significantly higher than in the E. coli-positive infertile group (p<0.001). E. coli O6 was the most prevalent serogroup found in both groups. However, there was no significant difference in the frequency of different serogroups of E. coil between the two groups (p=0.55). Conclusion: Despite the higher prevalence of E. coli among fertile men, E. coli had more detrimental effects on semen parameters in infertile men. There was no significant difference in E. coli serogroups between the fertile and infertile groups.

• Korean Journal of Animal Reproduction
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• v.26 no.1
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• pp.1-7
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• 2002

The present study was carried out to investigate the effects of maturation media on penetrability of pig oocytes by liquid boar sperm coincubated with different sperm concentrations in a modified Tris­buffered medium (mTBM). Follicular oocytes collected from ovaries of prepubertal gilts were matured in a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. The sperm­ich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ far 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes in 5% $CO_2$in air at 38.5$^{\circ}C$, cumulus cells were removed and oocytes (30~40) were coincubated for 6 h in 0.5 $m\ell$ mTBM fertilization medium with five different (1$\times$10$^{6}$ , 2$\times$10$^{6}$ , 4$\times$10$^{6}$ , 6$\times$10$^{6}$, 10$\times$10$^{6}$ $m\ell$) sperm concentrations. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ NCSU-23 culture medium fur further culture of 6 h. At 12 h after IVF, sperm penetration, polyspermy and male pronuclear formation of oocytes were evaluated. Oocytes of NCSU-23 maturation medium decreased polyspermy and increased male pronuclear formation compared to those of mTCM­199 and mWaymouth MB 752/1 maturation media. Of oocytes matured in NCSU-23 medium and inseminated in mTBM medium with 2~4$\times$10$^{6}$ $m\ell$ sperm concentrations, 50.8~50.9% showed sperm penetration, 13.3~19.5% polyspermy and 43.9~45.4% male pronuclear formation. In conclusion, we found out that oocytes matured in NCSU­23 medium and inseminated in mTBM medium showed superior in­vitro fertilization compared to those matured in mTCM­199 and mWaymouth MB 752/1 maturation media and inseminated in mTBM medium. The optimum sperm concentrations for in-vitro fertilization of oocytes matured in NCSU-23 medium by liquid boar semen stored at 17$^{\circ}C$ for 5 days were 2~4$\times$10$^{6}$ $m\ell$.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.1
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• pp.71-79
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• 1992

In vitro fertilization and embryo transfer (IVF & ET) is widely used for the males with subnormal or abnormal semen quality, as this was recommended in view of the relatively small numbers of spermatozoa required for fertilization and subsequent pregnancies could be obtained. The aim of this study is to know how the various functional parameters of spermatozoa in semen analysis affect the outcome IVF. This study was carried out between 1988-1989, with male factor patients selected on the basis of the semen quality. The selection criteria was based upon the mean values of concentration,% motility and % normal morphology from at least two semen analysis. There is a significant decrease in the fertilization and embryo transfer rates in the study group compared with control group (35.9% vs. 68% and 48.6% vs. 85.5% respectively), however, there was no significant difference in the pregnancy or delivery rates (19.6% vs. 21.4% and 60.0% vs. 62.5% respectively) per embryo transfer cycles. Fertilization rate is variously affected by the type and degree of sperm defect. No pregnancy was occurred in triple defect group and asthenoteratospermia group. There is no significant increase in the abortion rate in the male factor group. Improvement have to be made with the fertilization rate, as the pregnancy rate per OPU cycle in male factor group is still lower than that of normal group (9.5% vs. 18.3%). In conclusion, IVF can be used as a treatment for male factor infertility and the preparation of the semen sample can be modified to improve sperm recovery and obtain fertilization from abnormal semen samples.

• Journal of Embryo Transfer
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• v.23 no.3
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• pp.161-166
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• 2008

The techniques for the collection, cooling and freezing of semen and artificial insemination of horses are not fully understood in Korea. We investigated the percentages of total motile (TM) and progressively motile (PM) sperms after the collection, cooling and freezing of stallion semen. The average volume of semen was 167 ml in Thoroughbred and 68 ml in Arab. The average numbers of spermatozoa in Thoroughbred and Arab were $104\times10^6/ml$ and $86\times10^6/ml$ respectively. The average percentages of TM and PM were 82.3% and 88.6% in Thoroughbred, and 61.4% and 82.6% in Arab, respectively. The average percentage of TM at 4 hr after cooling at $5^{\circ}C$ was significantly lower than that at 0 hr ($30.0\pm4.1%\;vs.\;78.0\pm2.5%,\;p<0.05$), but the percentage of PM was similar between 66.5 and 73.2% at 0, 1, and 4hr. The average percentage of frozen-thawed Thoroughbred semen frozen in MFR5 extender was 56.2%, which was significantly higher than that of the semen frozen in LE extender (average 32.9%, p<0.05). The percentage of TM in Arab was similar for semen frozen in MFR5 extender and LE extender (18.2% and 21.2%, respectively), but the percentage of PM was significantly higher in sperm frozen in MFR5 extender than in sperm frozen in LE extender (69.0% vs. 36.4%, p<0.05). Four mares were artificially inseminated by Thoroughbred frozen-thawed semen and one of them fertilized at 11 day after artificial insemination. In this study, the collection, cooling and freezing of equine semen were possible under domestic conditions.

• Journal of Embryo Transfer
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• v.26 no.4
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• pp.245-250
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• 2011

The aim of this study was to evaluate the post-thawed characteristics of leopard cat semen. In this experiment, semen was collected from two leopard cats (A and B) at wild animal center in Seoul Grand Park in Korea. After collection, the sperms were washed with D-PBS and diluted by the freezing medium (Irvine science, USA) and stored in liquid nitrogen. The post-thawed concentration was $357{\times}10^6sperms/ml$ for A and $97{\times}10^6sperms/ml$ for B. The viability of post-thawed sperm from A and B individual was 24.0% and 19.0%, respectively. Pre-freezing motility of A and B individual semen was 68.54% and 56.65. Leopard cat A had more normal sperm than that of B (69.5% vs. 54.5%). Acrosome integrity analysis detected live (14.5% vs. 9.0%), damage (39.0% vs. 44.0%) and dead (46.0% vs. 47.0%) in leopard cat A and B, respectively. The present results concluded that leopard cat semen can be collected successfully by electro-ejaculation method and cryopreserved successfullyfor future use in different assisted reproductive technologies. The cryopreservation protocol needs to be modified for increasing post-thawed viability of leopard cat spermatozoa.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.2
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• pp.149-154
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• 2020

The aim of the current study was to compare the effects of season and breed on the reproductive performance of male and female sheep using 12 rams and 318 ewes of Assaf and Awassi breeds under the seasonal environmental condition of United Arab Emirates for two years. The blood level of testosterone hormone was measured monthly. Semen was collected twice a month from each male using artificial vagina and evaluated for volume, motility, livability, abnormality and concentration. The scrotal circumference and thickness as well as the left testicular length, width, height and volume were measured at one-month intervals. The level of testosterone in Assaf breed was significantly higher in autumn than winter and summer. The scrotal circumference and thickness as well as the left testicular length were significantly higher in Assaf breed than Awassi breed. While, left testicular width and volume were significantly higher in Awassi breed than Assaf breed. Scrotal circumference which was higher in spring and summer than in autumn and winter season in both breeds. The SCC of semen was significantly higher in autumn than in other seasons in both breeds. The sperm abnormality was significantly higher in summer than other seasons in both breeds. The livability was significantly lower in summer in both breeds. Fecundity and prolificacy were significantly higher in Assaf than Awassi breed during autumn season. Assaf breed showed the superior reproductive performance in the autumn season when compared with Awassi breed in the same season and other seasons. The Assaf breed tolerated the climatic conditions in UAE and kept the litter size of 1.72 in comparison to Awassi breed which showed litter size of 1.09. in conclusion, the results showed the superiority of Assaf over Awassi breed and offer a good model of breeding with increased fecundity and prolificacy specially in autumn season.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.2
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• pp.135-139
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• 2003

Objective: We investigated whether serum testosterone to estradiol ratio was decreased in infertile men and whether this condition can be corrected with oral aromatase inhibitor. Method: The serum testosterone to estradiol ratio of 26 men with testicular failure were compared with those of normal semen analysis parameter, 89 control reference group. All of 26 testicular failure group were diagnosed with the previous testicular biopsy. Then 46 men with oligospermia and/or asthenospermia were selected and treated with 1 mg of the aromatase inhibitor anastrozole ($Arimidex^{(R)}$) orally once daily for 3 months. Testosterone to estradiol ratio and semen analyses were evaluated during anastrozole therapy. Results: The testosterone level of testicular failure group was significantly lower and the testosterone to estradiol ratio was more decreased than normal semen parameter group. Forty six on-anastrozole group had significantly lower testosterone (4.6 versus 5.7 ng/ml, p<0.01) and higher estradiol (15.9 versus 23.4 pg/ml, p<0.01) than pre-anastrozole group, resulting in a decreased testosterone to estradiol ratio ($0.21{\pm}0.07$ versus $0.39{\pm}0.15$, p<0.01). Semen analyses before and during anastrozole treatment revealed significant increases in sperm count (35.5 versus 52.2 million sperm per ml, p<0.01) and motility (22.9% versus 29.3%, p<0.01). Conclusions: We identified infertile men with testicular failure had hormonal changes characterized by a decreased serum testosterone to estradiol ratio. The ratio can be corrected with aromatase inhibitor, resulting in a significant improvement in semen parameters.