• 한국임상수의학회지
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• 제13권2호
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• pp.114-122
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• 1996

The aim of this study was to assess the precision of the estimates of the time of estrous cycle, optimal breeding and ovulation derived by vaginal cytology. The thirteen Korea Jin-do dogs were examined the vaginal cytology, plasma estradiol-17$$\beta $ and progesterone assay during the estrous cycle. Day 0 was the day of the first male acceptance. The main change of vaginal cytology during the estrous cycle was the high proportion of anuclear cell and erythrocyte in proestrus, superficial cell, anuclear cell and erythrocyte in estrus, parabasal cell, large intermediate cell and leukocytes in diestrus, and parabasal cell and small intermediate cell in anestrus, respectively. These data indicated that vaginal cytology was reliable method for estimating estrous cycle in Korea Jin-do dogs. In the cell indices during estrus the maximum eosinoghilic index was $92.0{\pm}$2.6 (Mean{\pm} SEM$)% at Day 2 and the maximum cornification indez was $96.0{\pm}1.3%$ at Day 2, respectively. The eosinothilic indez and cornification indez of up to 70% were found at Day -1 to Day 5 and Day -6 to Day 8, and up to 80% at Day 1 to Day 4 and Day -4 to Day 6, respectively. From these data it was presumed that eosinophilic index was more reliable index for monitoring optimal breeding time than cornification indexm because eosinophilic index peak period was shorter than cornification indeX peak period and Day 2 was the day of ovulation. Therefore, optimal breeding time was the eosinophilic index peak period, more than 80% of eosinoghilic index. The $estradiol-17{\beta}$ peak, with 3 days delayed when progesterone concentration was $4.5{\pm}0.5 ng/ml$. These data estimated that the ovulation time was the day of eosinophilic index peak, Day 2. breeding time and pvulation time in Korea Jin-do dogs.

• 한국수정란이식학회지
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• 제27권2호
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• pp.93-100
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• 2012

To investigate the biochemical nature of changes in vaginal physiology during estrus and pregnancy, we examined the cytology and viscosity, and monitored the protein expression profile in vaginal mucus during estrus and pregnancy. The viscosity progressively decreased from estrus to pregnancy. Cell type analysis revealed that white blood cells progressively increased from estrus to pregnancy, while red blood cells progressively decreased during pregnancy. The cornification index (CI) was higher in estrus than in pregnancy. Protein mass spectrumetry identified the presence of ribosome-binding protein 1, GRIP 1 (Glutamate receptor-interacting protein 1)-associated protein 1, DUF729 (Domain of unknown function729) domain-containing protein 1, prolactin precursor, dihydrofolatereductase, and MMP (Matrix metalloprotease)-9 in vaginal mucus. MMP-2 and MMP-9 proteins in the vaginal mucus were active throughout estrus and gestation, as measured by a gelatinase assay, but most abundant in the vaginal mucus on day 0 of estrus. Results from ELISA of MMP-2 and MMP-9 were in accordance with the gelatinase assay. In light of the crucial role of metalloproteinases in extracellular matrix remodeling, the level of MMP-9 in vaginal mucus might be useful as an indicator of estrus and pregnancy to increase the efficiency of reproduction.

• 한국수정란이식학회지
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• 제23권1호
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• pp.13-18
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• 2008

Vaginal cytology with behavioral observation was performed in 34 estrous cycles in 16 Miniature Schnauzer dogs to evaluate the its usefulness. The mean duration of proestrus and estrus in Miniature Schnauzer based on behavioral observation and vaginal cytology was $7.58{\pm}1.50$ (Mean ${\pm}$S.D., range: $5{\sim}10$) and $8.46{\pm}2.82\;(6{\sim}17)$ days for proestrus, and $6.97{\pm}1.66\;(4{\sim}10)$ and $10.29{\pm}2.61\;(6{\sim}19)$ days for estrus, respectively. The duration of each phase of the estrous cycle was not significantly different based on between behavioral observation and vaginal cytology. The gestational length from the first day of male acceptance was $66.55{\pm}2.91\;(64{\sim}76)$ days, $57.70{\pm}1.92\;(54{\sim}62)$ days from the first male refusal, and $56.90{\pm}1.62\;(54{\sim}60)$ days from the onset of cytologic diestrus, respectively. Vaginal cytology during the estrous cycle were significantly characteristic of large intermediate cells in proestrus, anuclear cells in estrus, small intermediate cells in diestrus, and parabasal cells and small intermediate cells in anestrus (p<0.001), respectively. Cornification index (CI) by vaginal cytology was higher in proestrus ($69.90{\pm}1.44%$) and estrus ($91.35{\pm}1.09%$), then it was decreased in diestrus ($13.53{\pm}1.28%$) and anestrus ($1.16{\pm}1.26%$).

• 한국수정란이식학회지
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• 제28권1호
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• pp.25-30
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• 2013

The aim of this study was to estimate of ovulation time and parturition day at the same time as breeding in small dog by vaginal cytology and to confirm the accuracy by comparing the expected parturition day and the real one. 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, parabasal cell, small intermediate cell, large intermediate cell and leukocyte in diestrus, parabasal cell and small intermediate in anestrus, respectively. When day 0 was the parturition day, the period of pregnancy is 67.45(64~75) days when the cornification index (CI) is over 90%. Also, on the basis of ovulation day, 63.65(59~66) days was confirmed, and 57.0(52~60) days was confirmed based on the first day of diestrus. There are the gap of 4 days between the day being over 90% in CI and ovulation day. On the basis of this, when expecting parturition day based on the day being over 90% in CI by vaginal cytology, 18.1% was produced in the same of the expected parturition day and the real one, 30.3% and 33.3% were produced in the gap of one day and two days, respectively so, the accuracy within two days was 81.7%. In addition, based on the first day of diestrus, it also was identified to 81.7% as the difference between the expected parturition day and the real one within 2 days. It demonstrated there are any difference between any expected parturition day by vaginal cytology. Thus on the basis of the day of being over 90% CI, it is fully thought to using clinically due to the possibility of prediction the parturition day at the same time as the determination of the proper time of the optimal mating time.

• 한국임상수의학회지
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• 제36권1호
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• pp.38-41
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• 2019

The aim of this study was to estimate of prediction of whelping day and to confirm the accuracy of prediction of whelping day in small pet bitches. The gestation length from the each based on days was $66.64{\pm}2.32days$ ($Mean{\pm}S.D$) when the Cornification index (CI) was over 90% after the first vaginal discharge, $64.65{\pm}2.87days$ from the day of CI peak, $63.46{\pm}1.63days$ from the day of ovulation by progesterone concentrations, and $57.67{\pm}2.43days$ from the first day of cytologic diestrus, respectively. The whelping day was estimated 66 days when the CI was over 90% after the first vaginal discharge, 64 days from the day of CI peak, 63 days from the day of ovulation by progesterone concentrations, and 57 days from the first day of cytologic diestrus, respectively. The accuracy of the prediction of whelping day was 90.0% with a precision of ${\pm}2days$ when the CI was over 90% after the first vaginal discharge, 77.5% from the day of CI peak, 86.2% from the day of ovulation by progesterone concentrations, and 81.2% from the first day of cytologic diestrus, respectively. These results indicated that the prediction of parturition day by vaginal cytology was useful method for management of reproduction and parturition in small pet bitches.

• 한국수정란이식학회지
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• 제21권3호
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• pp.207-216
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• 2006

Shit-tzu견에서 발정 주기, 교배 적기 및 배란 시기를 정확하게 추정하기 위한 질 세포 검사를 확립하기 위하여 Shih-tzu 견 12두를 대상으로 발정 주기동안 질 세포 검사 및 estradiol-$17{\beta}$와 progesterone 농도를 측정하여 다음과 같은 결과를 얻었다. 발정주기별 질 세포상의 특징적인 변화로서 발정 전기에는 무핵 세포와 적혈구, 발정기에는 표층 세포와 무핵 세포 및 적혈구가 주종을 이루는 세포였으며 발정 휴지기에는 소형 중간 세포와 대형 중간 세포, 무발정기에는 부기저 세포와 소형 중간 세포 및 백혈구가 주종을 이루는 세포이었다. 또한 CI는 발정 전기와 발정기에 높게 나타났으나, 발정 휴지기와 무발정기에는 감소하여 각 기간별 비교에서 발정 전기와 발정기에 유의성 있게 높았다. 발정주기 중 첫 수컷 허용일(Day 0)을 기준으로 하였을 때 CI는 발정 전기 개시일 전후인 Day -9에 $72.4{\pm}2.9%$, Day -4에 $84.0{\pm}3.9%$로 80% 이상으로 증가하였으며 Day 0에 $97.0{\pm}1.1%$로 증가하여 Day 2에는 $98.2{\pm}0.5%$로 최고치를 나타내었으며 CI가 80% 이상을 나타낸 기간은 Day -4에서 Day 6까지(11일간)이었고, 90% 이상을 나타낸 기간은 Day -1에서 Day 5까지(7일간)이었다. 또한 임신 Shih-tzu 견 12두에서 혈장 progesterone 농도가 최초로 4.0 ng/ml 이상으로 증가한 날을 기준(Day 0)으로 발정 주기 중 혈장 progesterone과 estradiol-$17{\beta}$ 및 CI의 관계를 살펴보면 estradiol-$17{\beta}$가 Day -2에 최고치를 나타내었고, CI가 Day -1에 최고치를 나타낸 후 Day 0에 혈장 progesterone 농도가 $5.2{\pm}0.3ng/ml$로 최초로 4.0 ng/ml 이상으로 증가하였다. CI가 최초로 80 % 이상으로 증가하여 지속한 날은 estradiol-$17{\beta}$ peak 5일전인 Day -7이었고, Day -7부터 Day 3까지 11일 동안 80% 이상으로 지속되었다. CI가 최초로 90% 이상으로 증가한 날은 estradiol-$17{\beta}$ peak 2일전인 Day -4이었으며, Day -4부터 Day 2까지 7일 동안 90% 이상으로 지속되었다. 이상의 결과를 종합하여 보면 Shih-tzu 견에서 첫 수컷 허용일(Day 0)을 기준으로 하였을 때 CI 최고치는 Day 2에 나타났으며 이 때가 최상의 교배 적기로 사료되며, 혈장 progesterone농도가 4.0 ng/ml 이상으로 증가한 날(Bay 0)을 기준으로 하였을 때부터 CI는 혈장 estradiol-$17{\beta}$ peak 후 1일째인 최고치를 나타내었고, CI peak 후 1일째인 Day 0에 혈장 progesterone 농도가 최초로 4.0 ng/ml 이상으로 증가하여 CI가 90% 이상으로 지속된 시기가 최상의 교배 적기임이 확인되었다. 따라서 혈장 progesterone농도 측정으로 정확한 배란 시기 및 교배 적기를 판정할 수 있으나, 시설비가 저렴하고 검사 방법이 간단한 질 세포 검사가 Shih-tzu 견에서 발정 주기, 교배 적기 및 배란 시기의 판정에 응용될 수 있음을 시사하는 결과라고 사료된다.

• 한국임상수의학회지
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• 제19권4호
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• pp.418-425
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• 2002

Estrus was induced in 13 anestrus Korea Jin-do bitches by intramuscular injection of pregnant mare serum gonadotropin (PMSG) in a dose of 500 lU once daily for ten consecutive days, followed by an additional single intraveneous injection of 1,000 lU of human chorionic gonadotropin (hCG) on the tenth day. Day-changes of vaginal epithelial cells during the hormone treatment were investigated in each experimental bitches and compared with the those of spontaneous estrus bitches. The first days of vulval bleeding and male acceptance after PMSG treatment were on Day 6.0$\pm$ 1.5 (mean$\pm$ SD) and Day 9.0$\pm$ 1.9, respectively. And in all of 13 bitches, vulval swelling and perineal reflex were shown. The mean durations of proestrus and estrus were 2.9$\pm$ 1.4 (mean$\pm$ SD, range ; 1-6) and 11.5: 1.7 (range ; 8-14) days, respectively, that is, duration of proestrus was significantly shorter than that of the spontneous estrous bitches but duration of estrus was longer than that of the spontaneous estrous bitches. Characteristic features of vaginal cytology during the estrous cycle were the high proportions of large intermediate cell, superficial cell, anuclear cell and erythrocyte in proestrus, superficial cell and anuclear cell in estrus and parabasal cell, small intermediate, large intermediate cell, and leukocyte in diestrus, respectively. The comification index (Cl) was significantly high proportion in proestrus and estrus, when Day 0 was timed from the first day of male acceptance, the Cl was first increased above 80% on Day 0 and maintained above 80% until Day 0 to Day 5 during 6 days and showed a peak on Day 2. Also it was maintained above 90% until Day 2 to Day 3 during 2 days. These results indicated that all 13 ekperimental bitches showed positive estrus detection by the estrus behavior and vaginal smear test after treated with PMSG and hCC. It suggested that vaginal cytology was used to estimate the optimal mating and ovulation time, in consideration of the day when the Cl was maintained above 80% in estrus-induced Korea Jin-do bitches.

• 한국수정란이식학회지
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• 제21권2호
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• pp.109-119
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• 2006

비글개 6두에서 11회(임신견 7두, 비 임신견 4두)의 발정 주기 및 임신 기간 동안 질 상피 세포 검사 및 혈장 progesterone과 estradiol-$17{\beta}$ 농도를 측정하여 질 상피 세포상과 번식 호르몬의 관계를 조사하고 배란 및 교배 적기 판정을 위한 기초 자료를 제공하고자 본 실험을 실시하였다. 임신 예와 비 임신 예에 있어서 발정 전기, 발정기 및 발정 휴지기의 기간은 각각 $8.5{\pm}1.4(Mean{\pm}SD),\;10.0{\pm}1.4$ 및 $54.0{\pm}2.8$ 그리고 $7.9{\pm}2.1,\;9.5{\pm}0.7$ 및 $62.0{\pm}11.3$일이었다. 임선 예에 비해 비 임신견의 발정 휴지기가 길었으며, 발정 주기 사이의 간격은 임선 예가 $246.2{\pm}24.5$일, 비 임신 예은 $175.3{\pm}34.5$일을 나타내어 임신이 되었던 예가 임신이 되지 않았던 예보다 길었다. 발정 주기별 질 상피 세포상의 변화를 보면 발정 전기와 발정기에는 superficial cell, anuclear cell 및 적혈구가, 발정 휴지기에는 parabasal cell, small intermediate cell, large intermediate cell 및 백혈구가, 그리고 무발정기에는 parabasal cell과 small intermediate cell이 주종을 이루었다. Cornification index(CI)는 발정 주기와 발정기에 유의하게 높았다. 발정 주기 중 혈장 progesterone과 estradiol-$17{\beta}$의 농도 사이의 관계를 혈장 progesterone 농도가 최초로 4.0 ng/ml 이상으로 상승한 날을 기준(Day 0)으로 살펴보면, 혈장 progesterone 농도는 발정 출혈 개시 일에 1.0 ng/ml 이하였으나 Day -2에 2.0 ng/ml 이상으로 상승하였으며, 혈장 progesterone 농도가 최초로 4.0 ng/ml 이상으로 상승한 날은 최초 교배 허용 후 2일이었다. Day 20일 경에 임신 예 및 비임신 예 모두 40 ng/ml 이상으로 최고치를 나타내었으며 Day 35 일까지 임신 예와 비 임신 예 사이에 차이가 인정되지 않았으나 이후 임신 예가 비 임신 예에 비해 높게 유지된 기간이 걸었다. 혈장 estradiol-$17{\beta}$ 농도는 발정 출혈 개시 일에 9 pg/ml 이상을 나타냈으며 이후 급증하여 Day -2 에 26.4 pg/ml로 peak를 나타내었다. CI와 혈장 progesterone 및 estradiol-$17{\beta}$ 농도 사이의 관계를 살펴보면, 혈장 estradiol-$17{\beta}$ 농도는 최초 교배 허용 일(Day 0)에 peak를 나타냈고 혈장 progesterone 농도는 Day 2에 4.0 ng/ml 이상으로 처음 상승하였다. CI 는 혈장 estradiol-$17{\beta}$ peak 이후 l일(Day 1)에 최고치를 나타냈으며 80% 및 90% 이상으로 증가한 날은 각각 Day -1 및 Day 1이었다. CI가 80% 및 90% 이상을 지속한 기간은 각각 Day -1에서 Day 8(10 일간 ) 및 Day 1에서 Day 6(6 일간)이었다. CI가 최고치를 나타낸 후 l일 (Day 2)에 혈장 progesterone 농도가 4.0 ng/ml 이상으로 처음 증가하였다. 결과를 종합하면 본 연구에서 비글개의 배란은 혈장 estradiol-$17{\beta}$ peak 이후 3일 및 혈장 progesterone 농도가 4.0 ng/ml 이상으로 처 음 상승한 날에 일어나며, 교배 적기는 CI가 90% 이상으로 증가한 후 2일 이후와 혈장 progesterone 농도가 $2{\sim}25\;ng/ml$인 시기로 생각된다.

• 한국수정란이식학회지
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• 제25권1호
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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.

• 한국임상수의학회지
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• 제17권1호
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• pp.225-233
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• 2000

The aim in this study was to obtain the basic data for estimation of optimal mating time and ovulation time by finding out the relationship between vaginal cytology and reproductive hormone profiles during the estrous cycle in 36 Korea Jin-do bitches. A characteristic features of vaginal cytology during the estorous cycle were the high proportion of superficial cell, anuclear cell and erythrocyte in proestrus, superficial cell, anuclear cell and erythroucyte in estrus, parabasal cell, small and large intermediate cell and leukocytes in diestrus, and parabasal cell and small intermediate cell in anestrus, respectively. Cornification index(CI) was the high proportion in proestrus and estrus, then it was decreaed in diestrus and anestrus. It indicated that the CI was significantly high proportion in proestrus and estrus in comparison with the other phases. Plasma progesterone concentration was below 1.0ng/ml at the first day of vulval bleeding in pregnant and non-pregnat bitches, and then it was increaed slowly. When Day 0 was timed from the day that plasma progesterone concentration was first increased above 4.0ng/ml, plasma progesterone concentrations at Day 0 in pregnant and non-pregnant above 4.0 ng/ml, plasma progesterone concentrations at Day 0 in pregnant and non-pregnant bitches were $5.2{\pm}0.3ng/ml and 5.7{\pm}0.5ng/ml$$46.5{\pm}3.3 ng/ml$ in pregnant bitches and at Day 20 with $39.8{\pm}0.1ng/ml$ in nonpregnant bitches. It indicated that plasma progesterone concentration was higher in pregnant bitches than in non-pregnant bitches. Hereafter plasma progesterone concentration was decreased steadily. At Day 63 which is parturition day, plasma progesterone concentration was decreased below 1.0ng/ml with $0.8{\pm}0.2ng/ml$ in pregnant bitches whereas in the non-pregnant bitches at Day 75 were decreased below 1.0ng/ml with 40.5{\pm}0.4ng/ml$. Plasma progesterone concentrations was maintained below 1.0ng/ml during anestrus in all of them. The plasma estradiol-17 $\beta$ concentration was increased above 1.0 pg/ml at the first day of vulval bleeding and it showed a peak Day-1 with 38.2 pg/ml. Thereafter it was sharply decreased after Day 0, which was the day that plasma progesterone concentration was first increase above 4.0ng/ml, and was maintained below basal levels. In relationship between CI and reproductive hormones, plasma estradiol-17 $\beta$concentration showed a peak at Day-3 and CI showed a peak at Day-1 which was the second day after plasma estradiol-17 $\beta$ peak, and plasma progesterone concentration was first increased above 4.0ng/ml at Day 0 which was the first day after CI peak. CI was first increased above 80% at Day-6 which was the third day before plasma estradio-17 $\beta$ peak and it was maintained above 80% between Days-6 and Day 3 during 10 days, and showed above 90% at Day-3 which was the day that plasma estradiol-17 $\beta$peak and was maintained above 90% between Day-3 and Day 3 during 7 days. In conclusion, ovulation in Koran Jin-do bitches occurred at the first day after CI peak, at the third day after plasma estradiol-17 $\beta$peak and the day when plasma progesterone concentration was first increased above 4.0 ng/ml. And it was estimated that the optimal mating time was the day when the CI was maintained above 90% and plasma progesterone concentration was between 3.0~8.0ng/ml. Therefore plasma progesterone concentration measurement was used for determination of an accurate ovulation time and the optimal mating time but also vaginal cytology, which is low-priced with equipment and is the simple examination method, was reliable method for estimating estrous cycle, optimal breeding time and ovulation time in Korea Jin-do bitches.