The objective of this study was to examine the effect of vitamin B (pantothenic acid, folic acid, and myo-inositol) that was supplemented to embryo culture medium on in vitro development of parthenogenetically activated (PA) pig embryos. Cumulus-oocyte complexes derived from slaughtered ovaries were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones (hCG and eCG) for the first 22 h and then further cultured in hormone-free medium for an additional 22 h. After maturation culture, metaphase II oocytes that extruded 1st polar body were electrically activated and treated with cytochalasin B for 4 h. Then, PA embryos were cultured for 7 days in a modified NCSU-23 that was supplemented with pantothenic acid, myo-inositol, or folic acid at different concentrations () according to the experimental design. Myo-inositol added to culture medium did not show any beneficial or inhibitory effects on embryo cleavage and blastocyst formation. However, pantothenic acid significantly inhibited blastocyst formation compared to control (no addition) (24% vs. 36%, p<0.05). Folic acid () significantly (p<0.05) increased blastocyst formation (56%) compared to control (41%). Our results demonstrated that in vitro development of PA embryos was significantly influenced by vitamin B and addition of folic acid to culture medium improved in vitro development of pig PA embryos.
This study was conducted to find out the effects of artificial shrinkage (AS) on post-thaw development of bovine embryos. The blastocoelic cavity of blastocyst was punctured to remove its fluid contents and then incubated in the holding medium (HM) for 10 min. The punctured and non-punctured (control) blastocysts were equilibrated in vitrification solution 1 (VS1; TCM-199+20% FBS+10% EG) for 5 min and vitrification solution 2 (VS2; TCM199+20% FBS+35% EG+5% PVP+0.5 M Sucrose) for 1 min and vitrified by direct dropping into the liquid nitrogen. Vitrified blastocysts (punctured and control) were thawed and cultured in vitro (12 hr) for studying survival and hatching rates. The levels of shrinkage were measured by the volume of the blastocyst during equilibration in VS1 (at 1, 3 and 5 min of equilibration) and VS2 (at 30 and 60 sec of equilibration) that was considering the volume of non-punctured blastocyst in HM as 100%. The levels of shrinkage were higher in punctured group (62.4, 64.6, 64.3% at 1, 3 and 5 min in VS1; 50.6 and 52.7% at 30 and 60 sec in VS2) than control group (84.8, 86.6, 86.4% at 1, 3 and 5 min in VS1; 72.1 and 68.8% at 30 and 60 sec in VS2), but within each group the levels of shrinkage were similar. The survival (90.9%) and hatching (50.0%) rates of vitrified blastocysts at 12 hr post-thaw were higher in punctured group than that in control group (76.9% and 0.0% respectively). We confirmed that vitrification solutions (VS1 and VS2) have no toxic effect on the survival of blastocysts because the survival rates of blastocysts exposed to VS1 and VS2 for 24 hr were similar between punctured and control groups (94.3 vs. 96.0%; p>0.05). In conclusion, the preliminary data show that AS of blastocyst may improve survival and hatching rate after thawing.
본 연구에서는 OPU를 통한 체내 유래 난자를 이용한 수정란 생산 시 1개월에서 6개월까지의 연구 기간에 따른 난포 생성수, 난자 회수율, 난자 등급율, 배반포 생성률을 분석하여 공란우의 활용 기간에 관하여 조사하였다. 1. 채란 기간에 따른 난포 생성수는 1개월에서 5개월까지의 난포 생성수는 , , , , 개로 유의적인 차이가 없었으나, 6개월째에는개로 급격하게 줄어든 것을 알 수 있었다. 2. 채란 기간에 따른 난자 회수개수는 1개월에서 3개월까지는
The increase in the total number of cows on farms, which breed Korean Native Cattle (KNC), is associated with many problems. In particular, the services per conception and calving interval have increased. In this study, we examined the effect of dietary supplementation with vitamin and mineral complex on the estrus rate, pregnancy rate, gestation length, and birth weight of KNC calves. Multiparous or primiparous KNC were divided into 3 groups with 40 heads per group. Experimental group 1 was administered a consisting of 35,000 IU vitamin A, 100 IU vitamin E, 200 mg -carotene, 200 mg Zn methionine, and 1.5 mg Se. Experimental group 2 was administered a vitamin complex (100 g/day) consisting of 100 IU vitamin E and 200 mg -carotene. In the case of multiparous KNC, the estrus detection rate in the control group was 90.0% and those in the experimental groups were 75.0% to 95.0%. However, the first-service pregnancy rate after parturition in the control group was 41.2%, which was significantly lower than that in the experimental groups (71.0% to 76.7%; p<0.05). The average duration of pregnancy in the group supplemented with the vitamin complex was days, which was similar to that in the case of the control group. The birth weight of calves from cows fed with vitamin complex was 25.3 to 27.0 kg, which was similar to that in the case of the control group (25.2 to 26.0 kg). In the case of primiparous KNC, no differences in the estrus rate, pregnancy rate, gestation length, or the birth weight of calves were noted between the groups. Thus, dietary supplementation vitamin and mineral complex have no effect on the reproductive efficiency in primiparous cows, but the pregnancy rate was observed to have increased in multiparous KNC with these supplements.
난포낭종은 소 번식 장애의 주요 원인 중의 하나이며, 다양한 유전자의 변화는 여러 세포와 조직 기능에 영향을 준다. 이러한 유전자 변화는 낭종성 난소에서도 나타날 수 있다. 이온 및 수송체와 관련된 유전자 변화가 한우의 난포낭종을 유발할 수 있을 것이라는 가설 하에 난포낭종성 난포에서 발현 변화를 보이는 유전자를 찾기 위하여 마이크로어레이 분석을 수행하였다. 마이크로어레이 분석 결과, 난포낭종성 난포에서 FGG와 LRP8이 증가하고, SLC44A4, S
Many studies propose that dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I) is associated with neurodegenerative disorders, such as Parkinson's disease and Huntington's disease. Mammalian mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I) consists of at least 46 different subunits. In contrast, the NDI1 gene of Saccharomyces cerevisiae is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. With a recombinant adeno-associated virus vector carrying the NDI1 gene (rAAV-NDI1) as the gene delivery method, we were able to attain high transduction efficiencies even in the human epithelial cervical cancer cells that are difficult to transfect by lipofection or calcium phosphate precipitation methods. Using a rAAV-NDI1, we demonstrated that the Ndi1 enzyme is successfully expressed in HeLa cells. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced HeLa cells were not affected by rotenone which is inhibitor of complex I, but was inhibited by flavone and antimycin A. The NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the NDI1 gene failed to survive. In particular, in the NDI1-transduced cells, the yeast enzyme becomes integrated into the human respiratory chain. It is concluded that the NDI1 gene provides a potentially useful tool for gene therapy of mitochondrial diseases caused by complex I deficiency.
The specimens from 32 aborted fetus and 274 aborted cows were collected in 168 farms of Chonnam province from 2005 to 2008 and were tested the brucellosis. The results obtained are summarized as follows. In the 32 aborted fetus, bovine brucellosis was detected in 12 heads (37.5%), bovine viral diarrhea-mucosal disease was detected in 7 heads (19%), ainovirus infection was detected in 1 head (3.1%), and multi-infection of BVD and brucellosis was not detected, respectively. In the 306 cases of aborted fetus and cows, bovine brucellosis was detected in 44 heads (14%). Status of abortion were confirmed in 63 farms (38%) out of 168 farms from June to August. From the point of raising scale, studies found that 128 farms (76%) out of all raised under 20 heads. The incidence of abortion by brucellosis was mainly showed in 30 heads (68.1%) about 151~250 days of gestation. In the result of the 18 farms survey, the causes of infections were detected movement of infected cattle in 5 farms (28%), unknown cause in 12 farms (67%), and recurrence in 1 farm (5%). The results of this study suggest to take an advantage of the prevention and fundamental research for bovine brucellosis in Chonnam province.
The relationships of scrotal circumference (SC) to semen characteristics and the conception rate (70 days-nonreturn rate) of artificial insemination in farm were studied with 137 heads of bull in Hanwoo. The average and range of SC were cm and 26.0~52.5 cm, respectively. Hanwoo bulls were classified with SC, divided into 34 cm below group, 34~39 cm group, and 39 cm over group. The 5,487 semen records of 43 heads of bull from July. 1. 2007 to June. 30. 2008. were used to determine the relationships between SC and semen characteristics. The semen concentration and total sperm number of each group were 11.18, 16.68, and , and 69.83, 101.64 and /ejaculate. The bulls with 34 cm or more SC were higher than the bulls with 34 cm below in semen concentration and total sperm number (p<0.01). But between SC and semen volume have no significant relationship (p>0.05). The 9,862 mating records of 44 farm with 137 heads of bull were used to determine the relationships between SC and conception rate. The conception rate of 1st artificial insemination were 73.31, 74.16, and 77.33 % in each group. Also SC was positively correlated with pregnancy rate (r=0.12, p=0.17). These results indicate that SC correlates positively with semen characteristics, and maybe with pregnancy rate in Hanwoo.
The conception rate of cow is a major factor in farm management. The environment of farm and management of cow are the best influencing factors on conception rate, and the fertility of bull is the second influencing factor. In Hanwoo bull, however, the informations limited to performance and carcass traits have been offered to Hanwoo farmer. Therefore, this study analysed the estimated relative conception rates (ERCR) for estimation of fertility of bulls, using the 8,892 mating data with 116 heads of prove bull to produce progeny. Mean of least square means of conception rate after first insemination was 50.95% in bull herds. On the standard of this mean, ERCRs after first insemination of each bull were analysed. Values ranged from -26.1% to +21.0%, the difference was 47.1%. Among 116 heads of bull analysed, KPN582 showed the highest ERCR as 21.0%, KPN550 (18.3%), KPN656 (16.7%), KPN632 (15.8%), KPN690 (14.9%) were gone behind, but KPN621 was the lowest as -26.1%, KPN680 (-21.3%), KPN674 (-16.2%), KPN569 (-15.9%), KPN699 (-14.9%) were succeeded. If ERCRs of Hanwoo bull will be offered to Hanwoo farmer, it will be worthwhile.
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 days (), and days after the first PMSG treatment. The duration of proestrus and estrus was days, and 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.
This study was carried out the normal serial ultrasonographic appearance of the postpartum uterine involution with small pet bitches. Postpartum changes in uterine shape, architecture, echogenicity and diameter were monitored with ultrasonography in small pet bitches (Miniature Schnauzer, Cocker Spaniel and Shih-tzu). In the Miniature Schnauzer bitches, uterine diameter of the placental sites and interplacental sites were decreased from at 1 day to at 7 day, and at 1 day to at 7 day postpartum. In the Cocker Spaniel, the placental sites and interplacental sites were decreased from at 1 day to at 7 day, and at 1 day to at 7 day postpartum. In the Shih-tzu, the placental sites and interplacental sites were decreased from at 1 day to at 7 day, and at 1 day to at 7 day postpartum. And uterine diameter was no statistically significant difference among bitches (p>0.05). At 67 days, the uterine diameter in Miniature Schnauzer bitches were 6~7 mm both placental and interplacental sites, and the uterine horns showed uniform hypoechoic, tubular structures without enlargement. In present study, the involution of the uterus was completed at 67 days after parturition in Miniature Schnauzer, and 65 days in Cocker Spaniel and the Shih-tzu. There were no significant differences of normal postpartum uterine involution between small pet bitches (p>0.05). In conclusion, the postpartum involution of small pet bitches appeared to be completed normally at 65~67 days after parturition and could be identified by gross findings such as vaginal discharges and ultrasonographic findings of uterine shape and echogenicity.
A necropsy of a primiparous, 4-year-old, Himalayan tahr (Hemitragus jemlahicus) was performed in September 8, 2009. Typical appearances of ovaries, oviducts, uterine horns, cervix and caruncles were well shown. Five cervical folds were present in the cervix. Cervical opening can be reached in the length of less than 15 cm from the entrance of vagina. Development of two follicles was found in the left ovary even though breeding season of this species naturally starts in late November. The reproductive organs of a primiparous, 3-year-old, Corriedale sheep showed that differences in the length and morphological appearance of cervix exist between two species. Comparative understanding of reproductive systems in Himalayan tahr and Corriedale sheep could help advancing assisted reproductive technologies in feral goats.