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Studies on the Freezing of Boar Semen II . In Vitro and In Vivo Fertilizing Capacity of Frozen Boar Spermatozoa (돼지정액의 동결에 관한 연구 II. 동결한 돼지정액의 체내, 체외수정능력)

  • Kim K. S.;Song H. B.
    • Journal of Embryo Transfer
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    • v.20 no.1
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    • pp.1-8
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    • 2005
  • This experiment was carried out to investigate the effects of saccharide in the lactose-egg yolk(LEY) extender for freezing of boar semen on the viability, normal acrosome, fertilizable of in vitro or in vivo oocyte after thawed. Normal acrosome post-thawed spermatozoa was higher when increasing of glucose concentration in LEY extender with 3 or $4\%$ glycerol, but viability was not significant. Viability of the post-thawed spermatozoa was higher when fructose or fructose and glucose were added to LEY extender with $3\%$ glycerol than glucose and sucrose or fructose, glucose and sucrose(P<0.05). Rate of normal acrosome of post thawed spermatozoa was higher when both fructose and glucose$(81.4{\pm}2.3\%)$ were added to the LEY extender than saccharide not added$(41.6\pm0.6\%)$ to it(P<0.001). The percentage of fertilization, cleavage and development to blastocyst of oocytes fertilized with post-thawed spermatozoa from freezing by LEY extender were $70.8\~80.7\%$, $44.6\~45.7$ and $13.6\~16.0\%$, respectively. Conception rate by artificial insemination with frozen boa. semen was higher$(83.1{\pm}0.3\%)$ than commercial frozen semen from SGI company$(50.0{\pm}0.1\%,\;P<0.05)$, but litter size were no significant differences between frozen by LEY extender$(9.4{\pm}1.7\~10.4{\pm}0.7head/sow)$ and SGI semen$(8.0{\pm}1.1 head/sow)$.

Relationship between the Conception Rate after Estrus Induction using $PGF_2{\alpha}$ and Other Parameters in Holstein Dairy Cows (젖소에서 $PGF_2{\alpha}$ 투여에 의한 발정 유도 후 수태율과 다른 인자와의 관계)

  • Park, Chul-Ho;Lim, Won-Ho;Suh, Guk-Hyun;Oh, Ki-Seok;Son, Chang-Ho
    • Journal of Embryo Transfer
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    • v.25 no.3
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    • pp.133-139
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    • 2010
  • The purpose of this study was to determine the relationship between conception rate and other parameters (body condition score; BCS, progesterone concentrations and follicle size) before estrus induction with $PGF_2{\alpha}$. The conception rate in cows with (2.75, 2.75 to 3.25 and 3.25), BCS regardless of AI (artificial insemination) time was 47.5, 67.5% and 48.5% at $PGF_2{\alpha}$ injection, respectively. The conception rate regardless of BCS was 59.0% in cows inseminated based on detected estrus, and 46.2% in cows inseminated at 72 to 80 hours (timed artificial insemination, TAI) after $PGF_2{\alpha}$ injection. The conception rate regardless of AI time was 43.0% in cows with low progesterone concentrations (less than 1.0 ng/ml), and 67.5% in cows with high progesterone concentrations (more than 1.0 ng/ml) at $PGF_2{\alpha}$ injection. The conception rate regardless of progesterone concentrations was 59.9% in cows inseminated based on detected estrus, and 48.1% in cows of TAI after $PGF_2{\alpha}$ injection. The conception rate regardless of AI time was 36.0% in cows with small dominant follicles (less than 5 mm), 56.0% in cows between 5 mm to 10 mm of follicle size, and 65.5% in cows with large dominant follicles (more than 10 mm) at $PGF_2{\alpha}$ injection, respectively. The conception rate regardless of follicle size was 57.3% in cows inseminated based on detected estrus, and 47.6% in cows of TAI after $PGF_2{\alpha}$ injection. These results indicated that if the cows with BCS 2.75 to 3.25, active corpus luteum, and/or large dominant follicle (more than 10 mm) are used for estrus induction, the conception rate will be greater.

Confirmation of Estrus Induction using Vaginal Cytology in Shih-tzu Bitches (시츄견에서 질세포 검사를 이용한 발정 유기 효과의 확인)

  • Park, Chul-Ho;Lee, Suk-Kyung;Kim, Bang-Sil;Bae, Jae-Han;Ryu, Jae-Sun;Kim, Sang-Il;Lee, Ju-Hwan;Park, Sang-Guk;Park, In-Chul;Kim, Jong-Taek;Suh, Guk-Hyun;Oh, Ki-Seok;Son, Chang-Ho
    • Journal of Embryo Transfer
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    • v.25 no.1
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    • pp.67-75
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    • 2010
  • To investigate the change of vaginal epithelial cell in estrus-induced Shih-tzu bitch, estrus was induced by PMSG (50 IU/kg, for 10 days) and hCG (1,000 IU, on Day 10) in 12 anestrus Shih-tzu bitches. Day-changes of vaginal epithelial cells during the whole period of hormone treatment were investigated in each experimental bitches. The first day of vulvar bleeding and the first day of male acceptant was on $6.72{\pm}1.38$ days ($Mean{\pm}S.D.$), and $10.44{\pm}1.63$ days after the first PMSG treatment. The duration of proestrus and estrus was $4.36{\pm}1.20$ days, and $7.58{\pm}1.43$ days. Characteristic features of vaginal cytology during the estrous cycle were the high proportion of large intermediate cell, superficial cell, anuclear cell and erythrocyte in proestrus, superficial cell and anuclear cell in estrus, and parabasal cell, small and large intermediate cell, and leukocyte in diestrus, respectively. When it was timed from the first day of PMSG administration (Day 0), the cornification index (CI) was the high proportion in proestrus and estrus. The CI peak was maintained above 80% between Day 11~14 (4 days) and CI showed a peak at Day 12. These results indicated that the 12 Shih-tzu bitches showed positive estrus induction by vaginal smear test and observation of clinical estrus sign.

Ovarian response and surgical collection of embryos after induction of estrus in the bitch (발정유기견(發情誘起犬)의 난소반응(卵巢反應)과 수정란(受精卵)의 외과적(外科的) 회수(回收))

  • Kang, Byong-kyu;Choi, Han-sun;Na, Jin-su;Oh, Ki-seok;Son, Chang-ho;Lee, Cha-bum
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.373-381
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    • 1989
  • The aim of the present study was to test the efficiency of estrous induction in the premature, metestrous and anestrous bitches. The estrus was induced with prostaglandin $F_{2{\alpha}}$, estradiol-$17{\beta}$, pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin(HCG) in the treatment A, and with PMSG and HCG in the treatment B. Day 0 was the first day of estrone injection in the treatment A and the day of PMSG injection in the treatment B. Twenty three of the twenty six bitches were laparotomized under general anesthesia between 11 and 18 days after onset of behavioral estrus, whereas three bitches were not laparotomized and remained until parturition. Ovarian responses were evaluated with the total number of corpora lutea or ovulation sites. The uterine horns were flushed with phosphate-buffered saline added heat treated canine serum(10%), the flushing media was collected into watch glass and the ova were examined under stereomicroscope. The results obtained were as follows: 1. Standing estrus was observed on the day $17.7{\pm}1.5$ after injection of estrone in the treament A, but ovarian responses were not detectable. 2. Standing estrus was observed on the day $12.2{\pm}0.2$ after injection of PMSG in the treament Band 14 of 17 bitches showed ovarian responses. Ova were recovered in 9 of the 14 bitches. 3. Ovarian responses were observed in one of the three premature bitches. two of the three metestrous bitches and all of the 11 anestrous bitches. The average number of the ova collected from 9 bitchs were $12.2{\pm}1.4$. 4. Three bitches in the treament B exhibited behavioural estrus and all of them were mated with fertile male dog, resulting the pregnancy in only one bitch. The pregnant bitch gave the birth of two pups.

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Survey on Reproductive Traits of Average and High Yielding Holstein Cattle (젖소의 산유 능력에 따른 번식 성적 조사 연구)

  • Baek, K.S.;Lee, W.S.;Park, S.B.;Ahn, B.S.;Park, S.J.;Kim, H.S.;Kang, S.J.;Jeon, B.S.;Kim, S.B.;Son, J.K.
    • Journal of Embryo Transfer
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    • v.22 no.2
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    • pp.111-114
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    • 2007
  • This study was carried out to investigate the reproductive performance of average (less than 10,000 liters milk in 305 days) and high yielding (more than 10,000 liters milk in 305 days) Holstein cattle at commercial dairy herds (n=22). Data on milk progesterone (10 to 60 days postpartum), days to post-partum estrous, days to post-partum conception, service per conception and calving interval were recorded for two consecutive years. Post-partum milk progesterone concentration and days to reach peak milk progesterone concentration were similar in high and average yielding cows. High yielding cows took more days to show signs of first postpartum estrous than average yielding cows. Post-partum conception was 20 days earlier in average yielding cows than high yielding cows. Artificial insemination per conception was similar between average and high yielding cows. Calving interval was 26.9 days longer in high yielding cows compared to average yielding cows. In conclusion, better reproductive and feeding management may help improve the reproductive performance of high yielding dairy cattle in commercial dairy farms.

The Studies on In Vivo Embryo Production and Transfer in Hanwoo - II. Factors Influencing Pregnancy Rate after Embryo Transfer (한우에 있어서 체내수정란의 생산과 이식에 관한 연구 - II 한우 수정란 이식이 수태율에 미치는 요인)

  • 김덕임;서상원;정재경;이규승;서길웅;박창식;정영채;박병권
    • Journal of Embryo Transfer
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    • v.17 no.1
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    • pp.33-44
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    • 2002
  • These studies were carried out to establish an effective in vivo embryo transfer methods in Hanwoo by analyzing several factors that influence this process. In an embryo transfer, recipients with grade A corpus luteum of the right ovary and that of grade B of the left one showed a higher pregnancy rate(p<0.1) than others. The pregnancy rates of frozen embryos were significantly lower(P<0.01) than those of fresh ones; the former resulting in 35% and the latter resulting in 56.2%. Transfer of embryos according to the estrus cycle(6.0 ∼ 9.0 days) did not show a significant difference in pregnancy rate with fresh embryos recording 45.4 ∼65.7% and frozen ones recording 22.0 ∼ 50.0%. According to the status of corpus luteum and embryo freezing or not, the pregnancy rate was higher on grade A corpus luteum with 40.8 ∼67.9% than B and C which ranged from 25.0∼56.0%. The results of embryo transfer according to the development stage and grade of embryos showed that regardless of the embryo's grade. transfer of morula recorded an average pregnancy rate of 46.3%. This results higher than the transfer of blastocyst which was 34.1%.

재래산양의 과배란처리에 있어서 회수시간이 난자의 회수율에 미치는 영향

  • 박희성;홍승표;김선훈;정수영;진종인;김택석;이지삼
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.59-59
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    • 2003
  • 재래산양은 우리나라 고유의 유전자원으로서 보존뿐만 아니라 개량이 절실히 요구됨에도 불구하고 첨단생명공학기술을 이용한 연구가 극히 미진한 실정이다. 본 연구는 재래산양에 있어서 과배란처리에 의한 미수정 난자의 회수율 개선과 최적의 quality를 확보하기 위하여 과배란 처리 후 회수시간이 난포의 발달과 난자의 회수율에 미치는 영향을 조사하였다. 공시동물은 체중 15kg 전후의 성숙한 미경산 재래산양으로서 발정동기화를 위하여 CIDR(Eazi Breed, InterAg, New Zealand)를 10일간 질내에 삽입하고 과배란 처리는 FSH를 CIDR 삽입 8, 9, 10일째에 12시간 간격으로 70mg을 감량법으로 투여하였으며, PGF2$\alpha$는 8일째에 FSH와 함께 투여하고 CIDR제거는 10일째에 제거와 동시에 hCG 400IU를 투여하였다. 수정란회수는 외과적인 방법으로 실시하였으며, 난관에서 관류방법으로 난자를 회수한 다음 난포에서 흡입방법으로 난포란을 회수하였다. hCG 투여 후 29~34시간과 46~50시간에 관찰된 난포수는 두당 10.29개(144/14) 및 7.83개(47/6)였다. 또한 난포로부터 난포란의 채란율은 각각 66.0%(95/144) 및 53.2%(25/47)였으며, 두당 6.79$\pm$0.9개 및 4.17$\pm$1.1개의 난자를 채란하였다. hCG 투여 후 회수시간에 따른 난관에서 회수한 난자 수는 29~34시간 및 46~50시간 후 채란시 배란점은 평균 4.36개(61/14) 및 7.33개 (44/6)였으며, 난관에서 난자 채란율은 39.3%(24/61) 및 29.5%(13/44)였으며, 재래산양 한마리당 평균 1.71$\pm$0.5개 및 2.17$\pm$1.4개의 난자를 채란하였다. 난관으로부터 회수한 난자의 회수율은 29~34시간째에는 61개의 배란점을 확인하였으나 회수한 난자수는 24개로서 39.3%의 회수율을 보였고 두당 회수율은 1.71$\pm$0.5개였으며, 46~50시간에서는 29.5%의 회수율과 2.17$\pm$1.4개의 두당 회수율을 보였다. 회수한 난포란의 등급에 있어서 29~34시간에 등급별 회수율이 각각 GI(27.4%), GII(14.7%), GIII(43.2%), GIV(14.7%)로 나타났으며, 46~50시간에는 GI(12.0%), GII(12.0%), GIII(28.0%), GIV(48.0%)로 나타났다. 이상의 결과로 볼 때 미성숙 난자의 회수는 hCG 투여 후 29~34시간이 적합한 것으로 생각된다.

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Relationship between Estrous Expression, Body Condition Score (BCS) and Transferable Embryos Hanwoo Donors (한우 공란우의 발정발현율, 신체 충실 지수(BCS)와 이식가능 수정란과의 관계)

  • Son, Jun-Kyu;Choe, Chang-Yong;Cho, Sang-Rae;Lim, Hyun-Joo;Park, Seong-Jin;Baek, Kwang-Soo;Kim, Sang-Bum;Kwon, Eung-Gi;Jung, Yeon-Sub;Bok, Nan-Hee;Son, Dong-Soo
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.349-353
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    • 2011
  • 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.

Analysis of Estimation of Relative Conception Rate on Korean Proven Bull Number Semen using Non-return Rate (비발정 재귀율을 이용한 한우 보증씨수소 수정능력의 상대적 추정치 분석)

  • Yoon, Sung-Jae;Hwang, ChaeHyeon;Lee, Si-Hwa;Lee, Myeung-Sik;Lee, June-Sub;Rahman, Md. Saidur;Kwon, Woo-Sung;Park, Yoo-Jin;You, Young-Ah;Pang, Myung-Geol
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.213-217
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    • 2012
  • The decreased fertility is frequently thought to be problem of cattle production. However, studies figure out that number of these problems is related to bull factors especially in artificial insemination setting. Therefore, this study was designed to investigate the fertility status of bull by their estimated relative conception rate of cows that were inseminated by frozen semen from Korean proven bulls. Here we use the non-return rate (NRR) to access the bull fertility whereas, the NRR was define as the proportion of bulls that semen were used to inseminate cows and the number of cows that did not return for another service within 60 days. The data from 54,388 artificial inseminations (AI) were analyzed from 88 KPN semen. The NRRs of highest and lowest fertile bull were 83.81 and 51.33%, respectively. And mean NRR was 68.27%. In comparison to previously reported study, our data shows 17.38% higher NRR and the absolute value of difference in 50%>NRR and 50%

Relationship between Plasminogen Activator (PA) and HSP-90 in Uterus Tissue during the Porcine Estrous Cycle (돼지의 발정 주기 동안 자궁조직에서 Plasminogen Activator(PA)와 HSP-90과의 관계)

  • Gu, Ha-Na;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.219-223
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    • 2012
  • The present study was performed to identify the relationship between plasminogen activator (PA) and Heat Shock Protein-90 (HSP-90) in porcine uterus tissues during the estrous cycle. Porcine uterus tissues were obtained from preovulatory (Pre-Ov), post-ovulatory (Post-Ov) and early to mid-luteal (Early-mid L) stages. The protein was extracted from uterus tissue by using M-PER Mammalian Protein Extraction Reagent. Proteins were refined by RIPA Buffer and quantified by BCA methods. As results, t-PA expression was significantly (p<0.05) higher from pre-ovulatory(Epithelium tissue: $29,067{\mu}g/{\mu}l$, Myometrium tissue: $30,797{\mu}g/{\mu}l$) compared to the post-ovulatory stage(Epithelium tissue: $54,357{\mu}g/{\mu}l$, Myometrium tissue: $53,270{\mu}g/{\mu}l$) and early to mid-luteal stage(Epithelium tissue: $42,380{\mu}g/{\mu}l$, Myometrium tissue: $43,139{\mu}g/{\mu}l$). On the other hand, the uPA expression indicated higher from early to mid-luteal stage (Epithelium tissue: $0.02198{\mu}g/{\mu}l$, Myometrium tissue: $0.02412{\mu}g/{\mu}l$) than pre-ovulatory stage (Epithelium tissue: $0.01577{\mu}g/{\mu}l$, Myometrium tissue: $0.01531{\mu}g/{\mu}l$) and post-ovulatory stage(Epithelium tissue: $0.01414{\mu}g/{\mu}l$, Myometrium tissue: $0.01429{\mu}g/{\mu}l$). However, expression of u-PA did not differ from each estrous cycle in the epithelium tissue and myometrium tissue(p<0.05). Expression of HSP-90 was differ t-PA and u-PA from pre-ovulatory in Epithelium tissue($25,423{\mu}g/{\mu}l$) and early to mid-luteal stage in epithelium tissue($177,922{\mu}g/{\mu}l$) and myometrium tissue($26,664{\mu}g/{\mu}l$). These results suggest that HSP-90 and u-PA were related with change of uterus cycle according to the reformation of the tissues in porcine uterus.