The experiment was conducted to study the effect of GnRH administration at induced estrus on pituitary and ovarian response in buffalo heifers. Eight Murrah river buffaloes of 12 to 13 months of age were treated with $PGF_{{2}{\alpha}}$ and eCG combination. GnRH (Fertagyl) 200 ug was injected (iv) at estrus in four heifers (treated group) while saline (2 ml, iv) was injected in remaining four heifers (control group). Blood was collected through jugular catheter to estimate plasma FSH, LH and estradiol level. The pretreatment plasma FSH, LH and estradiol values ranged from $8.46{\pm}1.97ng/ml$ to $12.31{\pm}1.30ng/ml$, $0.87{\pm}0.21ng/ml$ to $1.19{\pm}0.29ng/ml$ and $19.09{\pm}2.38pg/ml$ to $20.24{\pm}1.00pg/ml$ respectively. The plasma estradiol concentration elevated significantly (p<0.05) within 24 hr after eCG administration and reached its peak levels of $154.09{\pm}17.28pg/ml$ and $181.95{\pm}31.82pg/ml$ at estrus in respectively treatment and control groups. The plasma FSH and LH concentrations did not increase during follicular development after eCG administration while initial significant (p<0.05) increases in both plasma FSH and LH concentrations occured within 5 and 10 min, reaching peak levels of respectively $110.06{\pm}23.56ng/ml$ and $13.15{\pm}3.13ng/ml$ within 90 min after GnRH injection was detected. A sharp and significant decline in plasma estradiol concentration ($59.27{\pm}8.78pg/ml$) associated with synchronized ovulation within 24 hours after GnRH injection was recorded. The observation suggest that the hypophysis of prepubertal buffaloes treated with eCG have gonadotrophins awaiting the releasing factor to evoke release of gonadotrophin during the follicular phase to induce synchronized ovulation.
Effective estrus detection and artificial insemination (AI) are necessary for profitable management of dairy herd. In current study, 45 crossbred lactating cows have been selected with the complaint of unobserved oestrus for more than sixty days postpartum. All cows had functional corpus luteum as examined by transrectal ultrasonography. Cows were treated with $PGF_2{\alpha}$ analogue and AI was performed with observed oestrus and then single dose of GnRH was administered. Similar synchronization protocol has been repeated after 14 days in cows that did not repose to first treatment. Remaining cows received additional $PGF_2{\alpha}$ after 14 days of second treatment and timed AI was performed following GnRH administration. Among 45 cows, 28.89% showed estrus after first treatment and 78.79% responded to second hormonal intervention. A higher conception rate (88.89% vs 26.66 and 72.72%) was observed in cows after triple administration of $PGF_2{\alpha}$ and timed AI. We noticed a significant differences in body condition score (BCS, 1~5 scale), postpartum period, and daily milk production between cows that either responded of non-responded following first and second hormonal treatment. In addition, there was a significant positive correlation between daily milk production and BCS, age and postpartum days, milk production and estrus/BCS, and milk production/BCS/estrus and conception rate. Depending upon the findings we conclude that hormonal intervention with $PGF_2{\alpha}$ and GnRH enhances postpartum ovarian cyclicity and help decreasing the days open of dairy herd. Therefore, this finding might provide an excellent guideline for target breeding system for profitable dairy herd management.
The objective of this study was to investigate the effects of abnormal ovarian cycles after superovulation treatment of Holstein Donor Cows. CIDRs were inserted into the vaginas of twenty two head of Holstein cows, regardless of estrous cycle. Superovulation was induced using folliclar stimulating hormone (FSH). For artificial insemination, donor cows were injected with $PGF_2{\alpha}$ and estrus was checked about 48 hours after the injection. Then they were treated with 4 straws of semen 3 times, with 12-hour intervals. Embryos were collected by a non-surgical method 7 days after the first artificial insemination. The cows were considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. Seventy two point seven percentage of the cows(16/22) had normal resumption of ovarian cyclicity(resumption within 40 days after superovulation), and 27.3%(6/22) 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 13.6%) were the most common types of delayed resumptions. The mean numbers of total ova from < 10 and $10{\leq}$ of corpora lutea(CL) was $7.8{\pm}1.8$ and $12.7{\pm}2.7$, respectively. The number of transferable embryos differed between < 10 and $10{\leq}$ CL was $5.4{\pm}1.3$ and $8.1{\pm}3.4$, respectively. Four point five percentage of the cows(1/22) did not resumption their ovarian cyclicity until 60 days after superovulation treatment. Diverse researches on the superovulation treatment method that is suitable for high-producing Holstein donor cows would contribute to preventing ovarian cyclicity disorder, as well as to the early multiplication of cows with superior genes by increasing the utilization value of donor cows.
The present study aimed at determining the effective dose of Folltropin, a follicle timulating hormone (FSH), on superovulation in indigenous cows of Bangladesh. Fifteen regularly cycling 5~7 years old dry cows, weighing 200~250 kg with 2.5~3.0 body condition scores (BCS) were divided into three groups (n=5). Individual groups were superovulated with 100, 200 or 300 mg of Folltropin per animal. The superovulation treatment was initiated at Day 10 or Day 11 of the estrous cycle (Day 0=day of estrus). Alfaprostol (6 mg) was injected to each cow 72 h after the initiation of superovulation treatment to induce eestrus. After confirming standing estrus, the cows were inseminated 2~3 times, 12 h apart, depending on the duration of estrus. At Day 6 or Day 7, individual horns of the uterus were flushed with 150~200 $m\ell$ of phosphate buffered saline supplemented with BSA (0.2%), penicillin (100 IU/$m\ell$) and streptomycin (100 $\mu\textrm{g}$/$m\ell$) using a two-way foley catheter. The embryos were concentrated, removing the excess medium through an embryo filter, and identified under a stereomicroscope. The identified embryos were collected, washed four times, evaluated and graded as excellent, good, fair or poor. The excellent, good and fair embryos were considered as transferable quality embryos. The mean (range). numbers of embryos collected vs. transferable quality embryos far 100, 200 and 300 mg of Folltropin were 4.5 (1~10) vs. 3.5 (1~8); 2.5 (1~4) vs. 1 (0~2) and 0.0 (0~0) vs. 0.0 (0~0), respectively, Folltropin at a dose of 100 or 200 mg produced suitable ovarian stimulation for superovulation in indigenous zebu cows of Bangladesh. A dose of 300 mg or more Folltropin consistently caused preovulatory corpora lutea formation in the ovaries and resulted in zero embryo recovery.
Ultrasonographic study was undertaken to establish the echogenecity and size of reproductive organs of 18 cycling buffaloes of different parities and compared with the sizes measured by palpation per rectum at estrus (day 0), met estrus (day 2), mid diestrus (day 10) and late diestrus (day 16). The overall mean size of cervix, uterine body, right horn, left horn, right ovary and left ovary measured by palpation per rectum were 2.70$\pm$0.43, 2.36$\pm$0.36, 2.17$\pm$0.37, 2.12$\pm$0.38, 2.63$\pm$0.41 and 2.72$\pm$0.37 cm, respectively. The corresponding ultrasonographic observations were 2.10$\pm$0.40, 1.85$\pm$0.30, 1.73$\pm$0.36, 1.64$\pm$0.37, 2.16$\pm$0.36 and 2.29$\pm$0.38 cm respectively. Variations in the size of genitalia due to stages of estrous cycle were non-significant. The size of genitalia measured by palpation per rectum was significantly higher (p<0.05) than by ultrasonography. However, there was linear positive correlation (r=+0.87) in the measurements by the two techniques. The ultrasonographic characteristics of tubular genitalia revealed different echogenic gray shades around the nonechogenic (black) central area of lumen depending upon the stage of cycle. The ovarian stroma appeared as hyperechoic (white) area with nonechogenic (black) follicle. The corpus luteum (CL) exhibited different echogenic texture viz. grayish black, grayish granular and grayish white at met estrus, mid diestrus and late diestrus, respectively. Therefore, ltrasonography can be effectively employed to record the exact size and echotexture of the buffalo genitalia during different stages of estrous cycle.
This study was carried out to establish an effective and practical system for commercialization of embryo production techniques by analyzing several factors influencing in vivo embryo production on days and seasons of flushing in Korean native cattle. In vivo embryos were flushed 226 times from 128 donors. The results obtained for the factors influencing in vivo embryo production on days and seasons of flushing were as follows :1.The percentages of fertilized, transferable and freezable embryos by seasons were significantly different in both FSR-P and SUPER-OV(P<0.01). The percentages of them were highest in sunrrner with FS H-P and highest in autumn with SUPER -OV.2. The production of transferable and freezable embryos by flushing days was highest in 8 days with FSH-P, and there was no difference between 7 and 8 days for SUP ER-OV. 3. The failure rates of recovery were 17.0% in SUPER-OV and 21.2% in FSH-P, respectively. The donors superovulated but failed recovery were 8.5% in SUPER-OV and 12.9% in FSH-P, respectively. Nonsuperovulated donors was 8.4% and donors giving less than 2 eggs at recovery was 8.4% in both FSH -P and SUPER-OV 4. The donors returned to normal estrus after superovulation were 34.1% after 1 cycle,39.4% after 2 cycles, and 16.7% after 3 cycles by FSH-P, respectively. For SUPER-OV, they were 55.3, 33.0 and 9.6%, respectively. Generally, normal estrus after the treatment of superovulation was earlier and the occurrence of ovarian cyst was also lower in SUP ER-OV than in FSH-P.5.The percentages of blastocyst in embryos flushed at 7~8 days after estrus were 21. 9% and 54.3% in FSH -P and SUPER-OV, respectively. The development of embryos was faster in SUPER-OV than in FSH-P.(Key words : in vivo embryo, flushing days, superovulation, FSH-P, SUPER-OV)
To establish the differential diagnosis and treatment method in bovine ovarian cysts, specially ovarian cysts with corpus luteum, serum progesterone concentration, rectal palpation and ultrasonography for measuring the cystic wall thickness and diameter of cyst and corpus luteum were investigated from 1,188 dairy cows with ovarian cysts. The plasma progesterone concentrations were 0.3$\pm$0.4 (mean$\pm$SD) ng/ml in 629 cows with follicular cysts, 3.7$\pm$1.1 ng/ml in 431 cows with luteal cysts, and 3.8$\pm$1.2 ng/ml in 128 cows with coexist of ovarian cysts and corpus luteum, respectively. The cystic wall thickness by ultrasonography were 1.6$\pm$0.4 mm in 629 cows with follicular cysts, 4.2$\pm$1.5 mm in 431 cows with luteal cysts, and 1.6$\pm$0.6 mm in 128 cows with coexist of ovarian cysts and corpus luteum, respectively. The days from initial treatment to insemination in follicular cysts were 28.1$\pm$6.9 days in treatment of GnRH alone, 15.9$\pm$2.9 days in combination of GnRH and dinoprost, and 15.1$\pm$3.1 days in combination of GnRH and cloprostenol. The percentages of cows conceived within 100 days after initial treatment were 61 %, 68% and 73% in treatment of GnRH alone, combination of GnRH and dinoprost, and combination of GnRH and cloprostenol, respectively. The days from initial treatment to insemination in luteal cysts were 3.8$\pm$0.6 days in treatment of dinoprost alone and 3.8$\pm$0.7 in cloprostenol alone. The percentages of cows conceived within 100 days after initial treatment were 69.5% and 68.5% in treatment of dinoprost and cloprostenol, respectively. The days from initial treatment to insemination in coexist of cysts and corpus luteum were 3.7$\pm$0.7 days in treatment of dinoprost alone and 3.8$\pm$0.6 in cloprostenol alone. The percentages of cows conceived within 100 days after initial treatment were 87% and 84% in treatment of dinoprost and cloprostenol, respectively. These results suggest that the best choice for treatment agents in ovarian cysts were combination of GnRH and PGF$_2$$\alpha$ in follicular cysts, and the PGF$_2$$\alpha$ in luteal cysts and in coexist of cysts and corpus luteum, respectively. In conclusion, it is suggest that ultrasonography is useful diagnostic tool for diagnosis and selection of treatment remedy in cystic ovaries of bovine.
본 연구는 한우에 PGF$_2$$\alpha$와 GnRH+PGF$_2$$\alpha$+GnRH(Ov-synch)를 처리하여 발정 및 배란을 동기화 시켰으며, 2차 GnRH 투여후 배란시간, 2차 GnRH 투여후 시간 경과에 따른 수태율과 발정ㆍ배란 동기화법에 의한 수태율을 조사하고자 실시하였다. 시험축은 총 4개 농가에서 경산우 150두를 무작위로 선발하여 시험에 공시하였으며, 발정ㆍ배란동기화 방법에 따라 발정을 유기한 후 1회 인공수정을 실시하고 수태율에 미치는 영향을 조사하였다. 호르몬처리 방법으로는 GnRH+PCF$_2$$\alpha$+GnRH(Ov-synch)와 PGF$_2$$\alpha$를 이용한 발정동기화 방법을 사용하였다. 2차 GnRH 투여 후 배란시간을 알아보기 GnRH 투여 후 24시간 후부터 32시간까지 2시간 간격으로 초음파 Sonovet-600(Medison. Korea)를 이용하여 난소를 촬영하였다. 1. 호르몬 투여후 발정동기 화율은 PGF$_2$$\alpha$ 투여구에서 40.0%와 GnRH+PGF$_2$$\alpha$+GnRH(Ov-synch)처리구에서 91.3%로 나타났다. 2. 2차 GnRH 주사후 24시간에 배란이 시작되어 32시간에 배란이 종료되었으며, 배란율은 28시간째에 46.6%가 배란되어 가장 높게 나타났다. 3. 2차 GnRH주사후 6∼24시간에 수정한 군이 6시간 이전과 30시간 이후에 수정시킨 군에 비해 높은 수태율을 나타냈다. 4. 호르몬 처리별 수태율은 PGF$_2$$\alpha$, CIDR 및 GnRH+PGF$_2$$\alpha$+GnRH(Ov-synch)에서 각각 50.0, 36.0와 76.9%로 GnRH+PGF$_2$$\alpha$+CnRH(Ov-synch)군에서 가장 높은 수태율을 나타냈다.
This research was investigated the relationship between the number of the transferable embryos and estrus expression rate, BCS (Body Condition Score), which affect the nutritional state of the cow, in Holstein donor cows. CIDRs were inserted into the vaginas of twenty two head of Holstein cows, regardless of estrous cycle. Superovulation was induced using folliclar stimulating hormone (FSH). For artificial insemination, donor cows were injected with $PGF_{2{\alpha}}$ and estrus was checked about 48 hours after the injection. Then they were treated with 4 straws of semen 3 times, with 12-hour intervals. Embryos were collected by a non-surgical method 7 days after the first artificial insemination. When BCS was $$\leq_-$$2.5, the total number of collected ova was 7.3 + 1.9, which is significantly lower (p<0.05) than the numbers 15.4 + 2.8 and 15.4 + 2.1 that were obtained when BCSs were 2.75 and $$\geq_-$$3.0, respectively. Whereas the numbers of transferable embryos were 5.2 + 1.4 when BCS was $$\leq_-$$2.5, which was smaller than the numbers 6.0 + 2.1 and 8.5 + 1.8 that were obtained when BCSs were 2.75 and $$\geq_-$$3.0, respectively; however, the differences were not significant. As for estrus induction rate, the cow groups whose BCSs were 2.75 and $$\geq_-$$3.0 showed 100.0% and 95.0%, respectively. Whereas the cow group whose BCS was $$\leq_-$$2.5 showed 57.1%, and the differences were significant (p< 0.05). As for estrous expression rate, the cow groups whose BCSs were $$\leq_-$$2.5, 2.75 and $$\geq_-$$3.0 showed 100.0%, 100.0% and 85.7%, respectively; however, the differences were not significant. According to the result of this research, it is considered that the total number of collected ova and the number of transferable embryos will be affected by the nutritional state before and after in vivo embryo production and superovulation treatment, and that although the mechanism is not clear, poor stockbreeding management and nutritional level would cause the decrease of ovum recovery rate and the number of transferable embryos in high-producing cows. On the other hand, diverse researches on the superovulation treatment method that is suitable for high-producing Holstein donor cows would contribute to preventing ovarian cyclicity disorder, as well as to the early multiplication of cows with superior genes by increasing the utilization value of donor cows.
The aim of this study was to evaluate the changes of protein patterns in granulosa cells and corpus luteum in ovaries during the estrus cycle in cows. The estrus cycle was devided into five steps of follicular, ovulatory, early-luteal, mid-luteal and late-luteal phases. In results, 61 spots of total 85 spots were repeated on follicular phase and 51 spots of total 114 spots were repeated on ovulatory phase. The 40 spots of total 129 spots were repeated on early-luteal phase and 49 spots of total 104 spots were repeated on mid-luteal phase. Also 41 spots of total 60 spots were repeated on late-luteal phase. On the other hands, the 16 spots were indicated difference in follicular phase and ovulation phase had a difference 10 spots. It was showed difference No. 103 spot in ovulation phase, No. 135 spot in early-luteal phase and No. 175 and 176 spots in mid-luteal phase. Also, the 11 spots were expressed specifically in mid-luteal phase and No. 178 and 179 spots were difference of expression in late-luteal phase. We confirmed that there were 7 spots for ovulation, 4 spots for luteinization and 2 spots for luteolysis. Spot No. 89~93 in ovulation phase were transferrin, and spot No.94~98 were HSP60. Spot No. 103 was Dusty PK, spot No. 135 was OGDC-E2, and spot No. 175 and 176 were Rab GDI beta from luteinization. Spot No. 178 and 179 in luteolysis were vimentin. This results suggest that will be help to basic data about infertility.
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