The present work describes experiments undertaken to evaluate the usefulness of selected physicochemical indices of semen, cell membrane integrity and sperm chromatin structure for the assessment of boar semen sensitivity to processes connected with pre-insemination procedures. The experiments were carried out on 30 boars: including 15 regarded as providers of sensitive semen and 15 regarded as providers of semen that is little sensitive to laboratory processing. The selection of boars for both groups was based on sperm morphology analyses, assuming secondary morphological change incidence in spermatozoa as the criterion. Two ejaculates were manually collected from each boar at an interval of 3 to 4 months. The following analyses were carried out for each ejaculate: sperm motility assessment, sperm pH measurement, sperm morphology assessment, sperm chromatin structure evaluation and cell membrane integrity assessment. The analyses were performed three times. Semen storage did not cause an increase in the incidence of secondary morphological changes in the group of boars considered to provide sperm of low sensitivity. On the other hand, with continued storage there was a marked increase in the incidence of spermatozoa with secondary morphological changes in the group of boars regarded as producing more sensitive semen. Ejaculates of group I boars evaluated directly after collection had an approximately 6% smaller share of spermatozoa with undamaged cell membranes than the ejaculates of boars in group II ($p{\leq}0.05$). In the process of time the percentage of spermatozoa with undamaged cell membranes decreased. The sperm of group I boars was characterised with a lower sperm motility than the semen of group II boars. After 1 hour of storing diluted semen, the sperm motility of boars producing highly sensitive semen was already 4% lower ($p{\leq}0.05$), and after 24 hours of storage it was 6.33% lower than that of the boars that produced semen with a low sensitivity. Factors that confirm the accuracy of insemination male selection can include a low rate of sperm motility decrease during the storage of diluted semen, low and contained incidence of secondary morphological changes in spermatozoa during semen storage and a high frequency of spermatozoa with undamaged cell membranes.
The objective of this study was to investigate the effects of abnormal ovarian cycles after superovulation treatment of Hanwoo donors. Thirty six, at random stages of the estrous cycle, received a CIDR. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU=1 ml) administered twice daily in constant doses over 4 days. On the 3th administration of FSH, CIDR was withdrawn and 25 mg $PGF_2{\alpha}$ was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received $100\;{\mu}g$ GnRH at the time of Ind insemination. Embryos were recovered 7 or 8 days after the 1st insemination. The cows were considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. 50.0 percentage of the cows (18/36) had normal resumption of ovarian cyclicity (resumption within 40 days after superovulation), and 50.0% (18/36) had delayed resumption(resumption did not occur until>40 days after superovulation). Delayed resumption Type II (first ovulation did not occur until $\geq$ 40 days after superovulation, i.e. delayed first ovulation 33.3%) were the most common types of delayed resumptions. The mean numbers of total ova from < 10 and 10$\leq$ of corpora lutea (CL) was 7.3 and 13.9, respectively. The number of transferable embryos differed between < 10 and 10$\leq$ CL was 4.2 and 5.1, respectively. 11.1 percentage of the cows (4/36) did not resumption their ovarian cyclicity until 60 days after superovulation treatment.
To evaluate the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 15 boars(Duroc, Landrace, and Yorkshire) and 2 mixed dogs which had been proved to be fertile in the past then, the semen were preserved in BWW medium at $4^{\circ}C$ or $18^{\circ}C$ for about 20 hours and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of sperm binding to the ova, penetration and formation of a male pronucleus, and the numbers of both bound and penetrated sperm per ovum. Both the semen preserved at $18^{\circ}C$ for about 20 hours and that treated by swim up procedure showed considerably higher rates of sperm binding and penetration as well as higher number of penetrated sperm than that preserved at $4^{\circ}C$ for about 20 hours, respectively(p<0.01). Motility of boar sperm at insemination was from 40 to 90% and no difference in hamster test was obtained according to different degree of sperm motility. Abnormality in morphology of boar sperm at insemination was from 6 to 45% and no difference in hamster test was obtained according to different degree of sperm abnormality. The sperm concentrations of $7{\times}10^7$ and $7{\times}10^6$ showed considerably higher rates of sperm binding and penetration as well as higher number of bound sperm than that of $7{\times}10^4$ (p<0.01) along with the same higher results than that of $7{\times}10^5$(0<0.05), respectively. Boar sperm showed considerably higher rates of sperm binding and penetration as well as higher numbers of both bound and penetrated sperm than dog sperm, when both semen were treated by BWW+heparin medium and swim up procedure, respectively. These results indicated that fertile boar sperm showed considerably lower rates in the results of hamster test, when preserved at $4^{\circ}C$ for about 20 hours and in lower concentration of sperm than when preserved at $18^{\circ}C$ for about 20 hours and in higher concentration of sperm, respectively, and at the same time considerably higher results than fertile dog sperm, consequently to prove that hamster test would be of great value in assaying the fertilizing capacity of boar sperm.
The objective of this study was investigate the superovulation treatment and to relate concentrations of blood urea nitrogen(BUN) in Hanwoo donors. Thirty six, at random stages of the estrous cycle, received a CIDR. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU=1 ml) administered twice daily in constant doses over 4 days. On the 3th administration of FSH, CIDR was withdrawn and 25 mg $PGF_2a$ was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received $100\;{\mu}g$ GnRH at the time of 1st insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Cows with BUN <10, 11~18 and ${\geq}$19 mg/dl had return of estrus of 34.6, 30.5 and 30.4 days respectively. Return of estrus after superovulation treatment was not significantly lower for cows with blood urea nitrogen (BUN) above 10 mg/dl than for cows with BUN below 10 mg/dl. Cows with BUN <10, 11~18 and ${\geq}$19 mg/dl had number of transferable embryos of $3.2{\pm}1.2$, $5.4{\pm}1.9$ and $4.1{\pm}2.1$ respectively.
The objective of this study was to investigate the relationship between estrous expression, body condition score (BCS), blood urea nitrogen (BUN) and number of transferable embryos for the purpose of improving reproductive performance in blood of Hanwoo donors. Sixty, at random stages of the estrous cycle, received a CIDR. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU=1 ml) administered twice daily in constant doses over 4 days. On the 3th administration of FSH, CIDR was withdrawn and 25 mg PGF2 ${\alpha}$ was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received 100 ${\mu}g$ GnRH at the time of 1nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. The estrous inducement rate and estrous expression rate were significantly lower for cows with BCS below 2.25 than for cows with BCS above 2.25. There was 50.0% of rate of mounting in cows with BCS below 2.25 whereas the rate of mounting was markedly increased in cows with BCS above 2.25 (94.1% and 89.5% for BCS 2.25~2.75 and BCS above 2.75 cows, respectively). Cows with BCS <2.25, 2.25~2.75 and ${\geq}$2.75 had number of transferable embryos of $4.5{\pm}0.7$, $5.9{\pm}1.8$ and $5.6{\pm}2.3$ respectively.
The objective of this study was to investigate the relationship between concentration of urea nitrogen, glucose, cholesterol and number of transferable embryos for the purpose of improving reproductive performance in blood of Hanwoo donors. Fifty five, at random stages of the estrous cycle, received a CIDR. Four days later the animals were superovulated with a total of 28AU FSH (Antorin, 2AU=1 ml) administered twice daily in constant doses over 4 days. On the 3th administration of FSH, CIDR was withdrawn and 25 mg $PGF_2$${\alpha}$ was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received 100 ${\mu}g$ GnRH at the time of 1nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Cows with BUN < 10, 11~18 and ${\geq}$19 mg/dl had number of transferable embryos of $4.32{\pm}1.3$, $5.8{\pm}1.8$ and $4.7{\pm}2.1$ respectively. The mean numbers of total ova from < 10 and 10${\leq}$ of corpora lutea(CL) was 8.9 and 14.3, respectively. The number of transferable embryos differed between < 10${\leq}$ CL was 4.8 and 5.6, respectively.
Park, Joung-Jun;Yoo, Han-Jun;Kim, Ki-Won;Lee, Seung-Hwan;Park, Choon-Keun;Hong, Seong-Koo
Reproductive and Developmental Biology
/
v.35
no.3
/
pp.233-238
/
2011
This study was performed to investigate the FSH levels for superovulation procedure in Korean Native Cattle (Hanwoo). The effectiveness of 200 mg and 400 mg of FSH to initiate superovulation was examined in Hanwoo. Donors, at random stages of the estrous cycle, received a CIDR 7 days later, 200 mg FSH group was treated with 40, 30, 20, 10 mg FSH levels in declining doses twice daily by intramuscular injection fur 4 days. Also, 400 mg FSH group was treated with 80, 60, 40, 20 mg FSH levels. On the 3rd day administration of FSH, 25 mg $PGF_2$${\alpha}$ was administered and CIDR was withdrawn. Donors were artificially inseminated twice at 12 hr intervals. The donor cattle received 250 ${\mu}g$ GnRH at time of 1st insemination and embryos were recovered 8 days after the 1st insemination. As a results, average number of CL treated with FSH 200 mg was higher as $20.9{\pm}1.20$ than $15.8{\pm}0.63$ for donors treated with FSH 400 mg, respectively(p<0.05). Treated group of 200 mg FSH level increased (p<0.05) the number of embryos recovered per procedure compared to 400 mg FSH level ($18.2{\pm}1.18$ vs. $12.38{\pm}0.52$, respectively). When treatment of 200 mg FSH was performed, average transferable embryos/ova increased (p<0.05) to $14.1{\pm}1.12$ from $6.8{\pm}0.33$ of treated of 400 mg FSH. Group of 200 mg FSH increased (p<0.05) to $8.3{\pm}0.76$ from $2.0{\pm}0.26$ in morula stage compare to 400 mg FSH group. Mean of total early blastocyst and expanded blastocyst stage embryos was similar (p<0.05) between the 200 mg and 400 mg FSH levels group ($4.7{\pm}1.19$ vs. $2.9{\pm}0.18$ and $1.2{\pm}0.40$ vs. $1.9{\pm}0.17$). These results suggest that 200 mg FSH level-based superovulation protocol with CIDR may be effectively used fur production of superior embryos in Hanwoo. In other words, the less level of FSH may be effectively applied for Hanwoo (Korean Native Cattle), because Hanwoo was smaller body size than beef or daily cow.
Park, Chul-Ho;Ryu, Jae-Sun;Yu, Dae-Jung;Park, In-Chul;Kim, Jong-Taek;Suh, Guk-Hyun;Oh, Ki-Seok;Son, Chang-Ho
Journal of Embryo Transfer
/
v.27
no.3
/
pp.133-139
/
2012
This study was carried out to develop the useful inducing method of estrus for Korean native cows. Under the condition of estrus induction by administering $PGF_{2{\alpha}}$ for the cows in which corpus luteum (CL) in ovaries was detected by ultrasonography, ovarian responses and the changes of progesterone ($P_4$) concentration against $PGF_{2{\alpha}}$ compared with conception rate were observed in cows and heifers. In inducing estrus administering $PGF_{2{\alpha}}$. to the cows which has corpus luteum in ovaries, ovarian reponses, the changes of progesterone concentration, and conception rate were identified and compared. The results attained from the studies were as follows. Significant decreases of CL in size over time after $PGF_{2{\alpha}}$ administration were detected in both cow and heifer groups (p<0.001), but not different between groups in the CL regression rate (p>0.05). In addition, the percentage changes relative to the plasma $P_4$ concentration on day 1 after $PGF_{2{\alpha}}$ treatment were decreased to below 1ng/ml. The growth rate of follicle was observed as 31% on day 1 and 42% on day 2 in cows, and 34% on day 1 and 97% on day 2 in heifers, resulting that growth of heifers are faster than that of cows (p<0.05). The conception rate after $PGF_{2{\alpha}}$ treatment were 60.5% and 64.2% in cows and heifers, respectively. It also indicated that the conception rate after estrus observation with $PGF_{2{\alpha}}$ injection was as high as 66.6% while that with timed-artificial insemination (TAI) regardless of the estrus observation was 56%, which means the pregnancy rate of artificial insemination after estrus observation was higher than that of TAI (p<0.05). In the result of all above, there were significant decreases in CL size and the plasma $P_4$ concentration by days but rapid growth in follicles, which has no differences in cows and heifers. The conception rate was commonly high after estrus observation and more than 50% under TAI.
The objective of this study was to investigate the relationship between body weight, body condition score (BCS), blood urea nitrogen (BUN), glucose, cholesterol and number of transferable embryos for the purpose of improving reproductive performance in Hanwoo donors. Seventy five cows, at random stages of the estrous cycle, received a CIDR together with injection of 1mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment begann. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU = 1 ml) administered twice daily in constant doses over 4 days. On the 3rd administration of FSH, CIDR was withdrawn and 25 mg $PGF_2{\alpha}$ was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received $100{\mu}g$ GnRH at the time of 1st insemination. Embryos were recovered 7 days after the 1st insemination. In conclusion, cows with body weight < 400, 400~450 and > 450kg had number of transferable embryos of $4.2{\pm}1.7$, $6.1{\pm}2.7$ and $4.8{\pm}2.6$, cows with BCS <2.25, 2.25~2.75 and ${\geq}2.75$ had number of transferable embryos of $4.6{\pm}1.6$, $5.7{\pm}2.4$ and $5.1{\pm}2.7$ respectively. These data indicate that a body weight and BCS for superovulation of CIDR-treated Korean native cows does not affect the embryo yield.
This study was designed to investigate the seasonal variation in semen characteristics and the change of motility during semen frozen/thawed, and conception rates were observed following AI at the different times after estrus synchronization. Semen collected from March to May showed significantly lower semen quality than the other months (P<0.05) and semen characteristics (volume, total sperm and motility) were significantly higher in October. Sperm motility after thawing in frozen semen were significantly lower in non-breeding season than in breeding season (P<0.05). Conception rate after treatment of estrus synchronization and AI different time after CIDR device removal, at 60 hour was higher than those of any other times through AI but there was no significantly difference between AI times. Semen characteristics change gradually during the breeding and non-breeding season. These results were considered as a model for the use of assisted reproductive techniques for AI of deer in Korea.
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