This study was carried out to investigate the semen characteristic, motility and viability and sperm motion characteristic by CASA test for establishing the Jindo-dog's semen freezing system. The results obtained are as follow: 1. The semen was collected 63 times. Average volume of semen, concentration of sperm, total number of sperm, progressive motility and viability were 3.8 $m\ell$ 145.6$\times$10$^{6}$ cells/$m\ell$, 396.2 x10$^{8}$ cells, 79.7% and 89.5%, respectively. Also, Fawn (Yellow) Jindo-dog comparing with White Jindo-dog showed better concentration of sperm, total number of sperm, progressive motility and viability. Among all dogs, the results of No. 2 Fawn Jindo-dog were the best. 2. The average progressive motility and viability of semen from 46 times were 73.5%, 82.3% before freezing and 51.1%, 64.9% after freezing. So, the freezing of semen has affected the progressive motility and viability. The progressive motility and viability of Fawn Jindo-dog's semen, before and after freezing, were better than White Jindo-dog. And No. 2 Fawn Jindo-dog showed the best results and showed significantly different among all dogs (P<0.05). 3. The 44 times-tested .esults by CASA system were as follow; MOT (motility) 65.6%. PROG (progressive motility) 54.8%, VAP (average path velocity) 75.3 ${\mu}{\textrm}{m}$/sec, VCL (curve linear velocity) 90.0 ${\mu}{\textrm}{m}$/sec, VSL (straight-line velocity) 69.4 ${\mu}{\textrm}{m}$/sec and ALH (amplitude of lateral head displacement) 4.4 ${\mu}{\textrm}{m}$. Although the motion characteristic of frozen semen were not significantly different between White and Fawn Jindo-dog, No. 2 Fawn Jindo-dog showed the best results and was significantly different among all dogs (P<0.05). 4. The success rate of frozen semen production between White and Fawn Jindo-dog were 43% (13/28), 94% (33/35), respectively, and the total success rate was 73% (46/63). The freezing-ability of Fawn Jindo-dog's semen was better than the other. Conclusively, the present results indicated that the characteristic and motility of Jindo-dog') semen were suitable for processing frozen semen, artificial insemination and mass production system. Also, the selection of suitable dog-breed was so important because the characteristic and freezing-ability of semen were significantly different between White and Fawn Jindo-dogs and among all individual dogs.
This study was conducted to determine the relationship between elapsed time after semen preservation on the changes of bacteria and semen quality. Semen was diluted with BTS(Beltsville Thawing Solution) extender without antibiotic for 7 days and sperm parameter and fertility were measured. Sperm motility was measured by CASA and total bacteria number was counted after 22~24 hr incubation from counting agar plate in which sperm dilute to $10{\sim}10^6$ in 0.9% saline solution and inoculate to agar. Acrosomal integrity was measured by Chlortetracycline (CTC) staining. CTC patterns were uniform fluorescence over the whole head (pattern F), characteristic of incapacitated acrosome-intact spermatozoa; fluorescence-free band in the post-acrosomal region (pattern B), characteristic of capacitated acrosome-intact spermatozoa; and almost no fluorescence over the whole head except for a thin band in the equatorial segment (pattern AR), characteristic of acrosome reacted spermatozoa. Total number of bacteria was significantly increased (p<0.0001) 3 days after preservation. Sperm motility, viability, and morphological abnormality on elapsed time after preservation were lower from 5 ($77.24{\pm}6.47$, p<0.001) and 7 days ($77.24{\pm}6.47$, p<0.001) after preservation compared to 1 ($15.71{\pm}7.18$) and 3 days($18.39{\pm}7.22$) after preservation, respectively. Sperm viability was significantly lower ($53.25{\pm}35.03$, p<0.0001) at 7 days after preservation. Morphological abnormality of sperm was lower (p<0.001) at 1 ($15.71{\pm}7.18$) and 3 ($18.39{\pm}7.22$) days compared to 5 ($21.84{\pm}7.91$) and 7 ($22.59{\pm}9.93$) days after preservation. Acrosomal integrity and capacitation rate (pattern F) were significantly lower (p<0.001) from 5 days after preservation. Based on the data we obtained from this study suggested that semen preserved more than 5 days without antibiotic would not recommend use for artificial insemination.
Oxidative stress is one of the major causes of failure of in vitro storage of boar semen. Reactive oxygen species (ROS) are one of the important mediators of oxidative stress during in vitro storage of boar semen. Our study examined the effects of taurine on sperm characteristic and on in vitro developmental embryos during in vitro storage of boar semen for 7 days. Semen was randomly aliquoted into 3 centrifuge tubes and treated with different concentrations of taurine (25-100 mM). The characteristics of boar sperm were analyzed for motility by light microscopy, viability by using a Makler counting chamber and membrane integrity by a hypoosmotic swelling test (HOST). The percentages of motile spermatozoa in taurine groups after 5 days were significantly higher compared to the control. Sperm viability in the control was lower than in taurine groups after 7 days irrespective of different taurine concentration. In the hyoosmotic swelling test (HOST), significantly higher results were obtained in taurine groups after 3 days. Also, the developmental rates of IVM/IVF porcine embryos from semen treated with pyruvate and taurine were significantly increased when compared with the control (p<0.05). These results indicate that supplementation of taurine as an antioxidant in boar semen extender can improve the semen quality.
Ji, Chang-Yoon;Kim, Tae-Ju;Lee, Seung-Ho;Kim, Young-Sik;Lee, Guem-San;Kim, Jung-Hoon;Choi, Goya;Ju, Young-Sung
The Journal of Korean Medicine
/
v.34
no.3
/
pp.1-12
/
2013
Objectives: This was study about identifying fine seed herbs that are highly mixed together: Celosiae Semen, Celoisae Cristatae Semen, Cuscutae Semen, Perillae Semen. Methods: In non-distinctive herbs through sensory test, we can find accurate discriminative points by using a stereoscope. As a result, a new discriminative key was completed. Results: We identified a comparison of herbs which are mixed up in distribution. 1) In the case of Celosiae Semen and Celoisae Cristatae Semen, Celoisae Cristatae Semen which is mixed up is bigger and more glossy and especially have the sunk umbilicus. 2) In the case of Cuscutae Semen and Perillae Semen, Perillae Semen is discriminated by characteristic morado netting surface and impression of fruit stalk in the single-ended. 3) In the case of Cuscutae Semen which is distributed in three species, we discriminated by beak shape, location of umbilicus and vomiting thread shape or not when put in water. (1) Cuscuta chinensis has a weak beaky shape, an umbilicus in center and is vomiting thread shape when put in water. (2) C. australis has very weak beaky shape, an umbilicus beneath and is vomiting thread shape when put in water. (3) C. japonica has clear beaky shape, an umbilicus beneath and is not vomiting thread shape when put in water. Conclusions: A stereoscope can be effectively used for identifying fine seed herbs hardly distinguishable by sensory tests.
This study was carried out to investigate the general characteristics such as semen volume, pH, sperm motility and sperm concentration of the semen collected from Korean Jindo dogs by the mothod of Digital manipulation of penis, and the effect of temperature and preservation time on motility of fresh semen. Multiple ejaculates were collected from four male Korean Jindo dogs. The results obtained in this experiment were as follows : 1. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the second fraction and the small volume of third fraction from the ejaculate were 3.29ml, 6.30, 96.70% and 1.64$\times$108 cells/ml, respectively. 2. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the first fraction from the ejaculate were 1.16ml, 6.10, 6.67% and 5.07$\times$105cells/ml. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the second fraction from the ejaculate were 2.30ml, 6.33, 97.66% and 1.92$\times$108cells/ml. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the third fraction from the ejaculate were 3.24ml, 6.51, 93.33% and 3.13$\times$107cells/ml. 3. Motility of fresh semen during preservation were higher at 17$^{\circ}C$ than at 5$^{\circ}C$ or 36$^{\circ}C$. When preservation temeprature was 17$^{\circ}C$, motility were 95.75% at 1 h, 90.00% at 6 h, 84.25% at 12 h, 68.00% at 18 h, 36.25% at 24 h and 28.75% at 30 h, respectively.
Preservation of liquid semen is an important factor for breeding management in swine industry. Oxidative stress of spermatozoa during liquid preservation has a detrimental effect on sperm quality and decreases fertility. Objective of this study was to determine the effect of antioxidant, Quercetin, on capability of porcine liquid semen preservation. Freshly collected porcine semen from boars (n=3), having proven fertility was counted, diluted to $3{\times}10^7/mL$ and divided into 5 different semen extenders. Aliquots of diluted semen with different extenders were subjected to measure the pH, motility, viability and sperm DNA structure status on elapse time after preservation for 10 days. For the first 3 days, semen preserved in all 5 different extenders maintained their initial pH and either gradually decreased or increased thereafter, indicating lipid peroxidation has started. Sperm motility (r=0.52, p=0.01) and viability (r=0.55, p=0.03) had positive correlation with semen pH. Sperm motility was maintained well (p<0.05) in especially 2 extenders containing Tris and antioxidant compared to other extenders, suggesting both Tris and antioxidant worked as pH regulator and had beneficial effects on sperm characteristic during preservation. Sperm DNA structure status accessed by sperm chromatin structure assay on elapsed time after preservation, tended to be higher in semen preserved without antioxidant. Taken together, addition of antioxidant to extender prevents the sperm from oxidative stress during storage in mechanism by which antioxidant slows the lipid peroxidation, and thus reduced the reactive oxygen species in preserved porcine semen resulted in maintaining semen pH, sperm motility and viability for 7~10 days.
Prostaglandin $F_2{\alpha}$ ($PGF_2{\alpha}$) can facilitate release of epinephrine from the adrenal gland. The objective was to extend these findings and determine the effects of $PGF_2{\alpha}$ on sexual activity and semen collection training in sexually inexperienced boars. Boars (n=32; $281{\pm}18$ days of age) were moved individually once weekly to a semen collection room equipped with an artificial sow. Before entering the semen collection room, boar received i.m. treatments of $PGF_2{\alpha}$ at doses of 5 (n=8), 10 (n=8), or 20 (n=8), and control boar (n=8) were not treated. Reaction time (elapsed time after entering collection pen until the start of mounting) for boars receiving 5mg ($3.3{\pm}0.9\;min$), 10mg ($3.3{\pm}0.8\;min$) $PGF_2{\alpha}$ was shorter (p<0.05) than for controls ($6.7{\pm}0.9min$). Duration of ejaculation (min) per session was longer (p<0.05) for $PGF_2{\alpha}$ (10 mg, 20 mg)-treated boars ($7.3{\pm}0.7\;min$, $6.9{\pm}0.7\;min$), compared to control ($3.4{\pm}0.8\;min$). The number of training session per boars was less (p=0.056) for $PGF_2{\alpha}$ 10mg-treated boars ($1.0{\pm}0.4$), compared to control ($2.0{\pm}0.4$). Semen characteristic such as volume, concentration, the number of total ejaculated sperm, were similar for $PGF_2{\alpha}$-treated and controls. There was no apparent difference on sperm movement characteristics (Mot: motility, VCL : curve linear velocity, VSL : straight line velocity, VAP : average path velocity, LIN : linearity) after semen preservation by collected with or without $PGF_2{\alpha}$ treatment. In summary, administration of $PGF_2{\alpha}$ in boars increased the sexual activity and facilitated the training boars to mount an artificial sow for semen collection, but did not affect semen characteristic.
Ji, D.Y.;Yoon, T.C.;Rho, J.R.;Cho, S.R.;Kim, C.K.;Pang, M.G.;Kim, Bo-Sook
Journal of Animal Science and Technology
/
v.49
no.5
/
pp.585-592
/
2007
This research was carried out in order to establish the production technique for Poong-san dog’s frozen semen, by examining the semen characteristic and the volume of glycerol added to the dilution solution, thawing temperature and sperm motility and viability as well as the motility using CASA according to time variation. Average semen volume was 5.9ml, sperm concentration 116.3×106 sperm/ml, total sperm number 789.3×106 sperm, motility 88.7±1.7% and viability 87.6±7.8%. When it was cryopreservation and thawed at different glycerol concentrated extender, it showed 52.7% motility and 57.7±10.3% viability at 7% glycerol, compared to other treatments. For semen cryogeny, at conditions of 5, 7cm and a height of 10cm for pre-cryogeny and maintaining the semen at 7cm from the surface of liquid nitrogen resulted in profitable motility and viability.
Kim, Kiju;Park, Soyeon;Cho, Youngjae;Jung, Bae-Dong;Park, Joung-Jun;Hahn, Tae-Wook
Korean Journal of Veterinary Research
/
v.54
no.4
/
pp.219-224
/
2014
Uterine sterilization is important for improving fertility in cattle. This study compared bacterial flora in the uterus between healthy and repeat breeder cows (RBCs). The uterine flushing of six heifers, 13 healthy HanWoo cows and eight RBCs (HanWoo) were sampled, and 15 frozen semen samples were selected. Overall, 35 bacteria were identified from in HanWoo uterine flushing and semen. The bacterial genera identified from HanWoo uterine flushing were Alloiococcus, Bacillus, Enterobacter, Enterococcus, Erysipelothrix, Gardnerella, Granulicatella, Kocuria, Pantoea, Pasteurella, Rothia, Serratia, Sphingomonas, Staphylococcus, Stenotrophomonas and Streptococcus. The bacterial genera identified from HanWoo semen were Bacillus, Escherichia, Kocuria, Oligella, Pseudomonas, Serratia, Sphingomonas, Staphylococcus, Stenotrophomonas and Streptococcus. The prevalence and presence of the identified bacteria between healthy cows and RBCs differed significantly. Further studies are needed to determine the role of these bacteria in the uterus of HanWoo cattle with reproductive disorder.
Moubasher, Alaa El din-Abdel Aal;Taha, Emad Abdelrehim;Elnashar, Ehab Mohamed;Maged, Ahmed Abdel Aal Abdel;Zahran, Asmaa Mohamed;Sayed, Heba Hassan;Gaber, Hisham Diab
Clinical and Experimental Reproductive Medicine
/
v.48
no.1
/
pp.61-68
/
2021
Objective: This study was conducted to investigate the relationship of semen parameters in samples used for intracytoplasmic sperm injection (ICSI) with fertilization and pregnancy rates in infertile couples. Methods: In this prospective study of Infertile couples with male factor infertility that had undergone ICSI, fractions of the same semen samples obtained for microinjection (to ensure the best predictability) were evaluated to determine the semen parameters and sperm DNA fragmentation index (DFI) on the day of oocyte recovery. Results: In total, 120 couples completed the study and were subdivided into fertilized (n=87) and non-fertilized couples (n=33). The fertilized couples were further classified into pregnant (n=48) and non-pregnant (n=39) couples. Compared to non-fertilized and non-pregnant couples, fertilized and pregnant couples showed statistically significantly higher sperm viability and percentage of normal sperm morphology, as well as significantly lower sperm DFI values. A receiver operating characteristic curve analysis of data from the 120 ICSI cycles showed that sperm viability, normal sperm morphology percentages, and sperm DFI were significant prognostic indicators of fertilization at cutoff values of 40%, 7%, and 46%, respectively. A sperm DFI of 46% showed sensitivity and specificity of 95% and 90%, respectively, for predicting fertilization, and no clinical pregnancies occurred in couples with a sperm DFI above 46%. Conclusion: Semen parameters from the ICSI day sample, especially sperm viability, normal morphology, and DFI, had an impact on fertilization and pregnancy outcomes in ICSI cycles.
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