A combined technology of transvaginal ovum pick-up(OPU) system with in vitro-oocyte manipulation technique can be used for improving reproductive efficiency in the cattle. The objective of this study was to establish a newly-conceived breeding program using OPU in the pregnant cows. The OPU trial was performed in pregnant cows every 10 days from 40 through 90 days of artificial insemination (Al), and number of follicles in ovary, number of retrieved oocytes and embryo development following in vitro-fertilization, were evaluated. Reduced number of follicles in the ovaries of pregnant cows was firstly detected from 70 days after A' and a significant (P<0.05) decrease in the follicle number (5.4 follicles /donor) was found at 90 days than at 40, 50, 60 and 80 days after Al (8.0~9.2). A similar pattern was also observed in the number of oocytes retrieved by OPU apparatus during experimental period. When retrieved oocytes were matured and inseminated in vitro with frozen bull semen, development of the oocytes to the blastocyst stage was not significantly affected by the retrieval time. Four embryos (morula or blastocyst stage) derived from oocytes retrieved from pregnant cows were nonsurgically transferred to four recipient cows on day 7 of estrus cycle. For the first time in Korea, three of four transferred embryos developed to live calves with normal physiological parameters. In conclusion, an effective breeding program employing pregnant cow can be developed by use of OPU trial and in vitro culture techniques of oocytes ; OPU system could be repeated in pregnant cows with no risk of abortion and viable offsprings were borne after transfer to the recipients.
The experiment was carried out to determine the concentrations of total calcium and inorganic phosphorus of thirty rera obtained from pregnant Holitein cows, six sera from normal non-pregnant cows with one month after delivery, six sera from cows with ovarian cyst and six sera from heifer of sixteen months old. The changes of total calcium and inorganic phosphorus were observed on the 2nd, 4th, 6th, 8th, and l0th month of gestation. The results obtained in this work were summarized as follows: 1. The mean values of total serum calcium obtained from the pregnant group were $9.59{\pm}0.34mg/100ml$, from the non-pregnant group $10.23{\pm}0.47mg/100ml$, from the infertility grou $10.37{\pm}0.27mg/100ml$, and from the heifer group $10.07{\pm}0.20mg/ml$. The change of total serum calcium values in the pregnant group revealed the tendency of decrease as gestation progresses and the lowest value appeared on the sixth month. The mean value of total serum calcium in the pregnant group was lower than those of the other groups, but the differences were not statistically significant. 2. The mean values of serum inorganic phosphorus obtained from the pregnant group were $5.82{\pm}0.31mg/100ml$, from the non-pregnant group $5.92{\pm}0.44mg/100ml$, from the infertility group $5.80{\pm}0.49mg/100ml$, and from the heifer group $5.10{\pm}0.32mg/100ml$. The change during pregnancy showed a slight fall and the lowest value appeared on the sixth month. The differences of serum inorganic phosphorus values between the pregnant group and the other adult groups were not significant, but these adult groups showed a significantly higher value than that of heifer group (p<0.05).
In an effort to confirm true oestrus and to detect early pregnancy in Zebu cows (Bos indicus), sequential blood and milk samples were collected at the day of imsemination (Day 0) and days 14, 20 and 24 after insemination. Progesterone was determined in skimmed milk and plasma by solid phase radioimmunoassay (RIA). Of the cows thought to be in oestrus plasma, (n = 46) and milk (n = 58) samples demonstrated low progesterone concentrations ($0.99{\pm}0.11$ and $2.02{\pm}0.14nmol/l$) in 42 (91%) and 52 (90%) cases respectively. Thirty two (76%) and 30 (71%) cows were thought to be pregnant by plasma progesterone RIA ($20.23{\pm}1.03$ and $20.48{\pm}1.01nmol/l$) at days 20 and 24 respectively. At the same period, 40 (77%) and 37 (71%) cows were thought to be pregnant by milk progesterone RIA ($27.82{\pm}1.28$and $28.02{\pm}1.27nmol/l$). Assuming 100% accuracy for rectal examination of pregnancy diagnosis between days 60-65 postservice, the RIA was found to be 84% and 90% accurate for plasma and 84% and 92% accurate for milk at day 20 and 24, respectively. All cows thought to be non pregnant by progesterone measurement were correctly diagnosed. Progesterone assay at 24 days after oestrus may therefore be accurate for early diagnosis of pregnancy in Zebu cows.
This study was carried out to investigate the effect, range of practice, and propriety for diagnosis of early non-pregnancies and reproductive disorders by dairy cows' milk progesterone analysis used EIA-kit of home products. The results were summarized as follows : 1. During 2 to 6 months after artificial insemination, the results of milk progesterone measurement by Home-kit and Auto ELISA reader-kit with pregnant dairy cows(152 heads) certified by rectal palpation were revealed, in Home-kit, 145 heads(95.4%) of positive reaction, 7 heads(4.6%) of quasi-positive, and 0 heads(0%) of negative among 152 heads and, in Auto ELISA reader-kit, 152 heads(100%) of positive reaction among 152 heads. 2. During 19 to 22 days after artificial insemination, the results of milk progesterone measurement by Home-kit, and thereafrer during 50 to 90 days after that, the results of pregnant test by rectal palpation were summarized as follows : 147 heads(82.1%) among 179 heads of positive reaction by Home-kit and 5 heads(31.3%) among 16 heads of quasi-positive were revealed pregnant cows by rectal palpation, and 42 heads(100%) among 42 heads of negative were non-pregnant. 3. During 19 to 22 days after artificial insemination, the results of milk progesterone measurement by Auto ELISA reader-kit, and thereafrer during 50 to 90 days after that, the results of pregnant test by rectal palpation were summarized as follows : 146 heads(86.9%) among 168 heads of positive reaction by Auto ELISA reader-kit and 6 heads(28.6%) among 21 heads of quasi-positive were revealed pregnant cows by rectal palpation, and 48 heads (100%) among 48 heads of negative were non-pregnant. 4. For the accuracy of the rectal palpation, Home-kit and Auto ELISA reader-kit were used in the cows of ovarian diseases. The results were following : in the cows of reproductive disorders expected negative milk progesterone, the accuracies of rectal palpation were the same 75.5%(40 heads among 53 heads) by Home-kit and Auto ELISA reader-kit, and in the cows of reproductive disorders expected positive milk progesterone, the accuracies of rectal palpation were 82.6%(19 heads among 23 heads) and 91.3%(21 heads among 23 heads) by Home-kit and Auto ELISA reader-kit, respectively, and the general accuracies of rectal palpation were 77.6%(59 heads among 76 heads) and 80.3%(61 heads among 76 heads) by Home-kit and Auto ELISA reader-kit, respectively.
These studies were performed to provide some basic informations for developing an automatic system in dairy farming cattle in order that the farmers may easily and automatically detect the maintenance of pregnancy and the fact of abortion of the pregnant cows and also to find out the diseased animals with fever. As a method of automatical detection of the maintenance of pregnancy or the fact of abortion, weighing the pregnant cows was conducted from one month-pregnancy to the term using a digital balance. From the first to the 3rd month of pregnancy the body weight of dairy cows was slowly increased (less than 2% per month), then, relatively high increase (3.4% -4.3% per month) from the fourth to the seventh month followed by decrease (3.3%) in the 8th month and very low increase (0.8-0.9%) from the 9th month to the term were shown, resulting in increase of 128.8 kg (25.05%) of body weight to be compared with the first weight. More than 107, increase of body weight to be compared with the first month-weight was denoted from the 61th month of pregnancy and more than 20% increase from the 7th or the 8th month of pregnancy as wells consequently it was presumed that detection of the maintenance of pregnancy is possible from the 4th or the 5th month of pregnancy. It was possible to diagnose a cow aborted at the 6th month by continual weighing the cow from the 1st month of pregnancy. The calved cows showed considerably higher decrease of body weight even in the third week after parturition (p<0.01)to be compared with the body weight near to the term (81.8-102.0 kg, 14-16% decrease). During the same period of 8months, the pregnant cows gained 127.4 kg (24.78% increase), whereas the non-pregnant cows gained 33.0 kg (0.71% increase) to be compared with the first weight showing considerably higher increase of body weight gain in the pregnant cows than the non-pregnant cows (p<0.01). The statistics of body temperatures of dairy cattle were collected from three clinics including the Teaching Hospital of Chonbuk University and the diseases were classified simply by the major symptoms manifested, denoting the highest temperature in respiratory disease ($39.8{\circ}C$) and the lowest in alimentary disease ($39.6{\circ} C$). These informations of body temperatures were expected to be of value for early and automatical detection of the diseased animals with fever when automatic machinery would be established. The results of periodic weighing the body weight of pregnant cows while milking were also expected to be of great use for the farmers to detect the maintenance of pregnancy and the fact of abortion when the automatic system is established in the near future.
Blood serum from 30 pregnant cows, 6 non-Pregnant cows (one month after delivery), 6 sterile cows (ovarian cyst), and 6 heifers (16 months old) were analyzed for total vitamin E concentration by the ferric chloride-bipyridyl reaction. The pregnant group was determined periodically throughout pregnancy. The results obtained were as follows: 1. The mean value of serum vitamin E level in pregnant group was $0.651{\pm}0.068(SE)mg/100ml$, in non-pregnant group $0.647{\pm}0.119(SE)mg/100ml$, in sterile group $0.488{\pm}0.053(SE)mg/100ml$, and in heifer group $0.083{\pm}0.016(SE)mg/100ml$. Samples below 0.19mg/100ml were net observed and most of the pregnants (83.3%) showed the values of 0.40 to 0.99mg/100ml during pregnancy 2. The difference of vitamin E level between pregnant group and non-pregnant group was net significant, but these trio groups showed a significantly higher value than that of sterile group. On the other hand, in heifer group was recognized significantly lower value compared with adult groups. 3. The change of serum vitamin E level in pregnant group revealed the tendency of decrease as gestation progresses and significantly dropped at the eighth month, thereafter, the value was rose slightly at the tenth month of gestation but no differences were observed. The coefficient of correlation between months and serum vitamin E levels during pregnancy was r=-0.956(p<0.01), and a regression equation was Y (serum vitamin E levels, mg/100ml)=1.034-0.063x(month).
In order to evaluate conception rate of Hanwoo in northwestern region of Gyeongsang-nam-do, we investigated conception rate and reduction of reproductive disorder rate after artificial insemination (AI) in 1,000 heads of breeding cows, This study showed that 80.9% of cows were classified as fertility after 1st and 2nd AI. For a accurate pregnancy diagnosis with practicing ovariectomy and histeotomy, we comparatively investigated each of 80 slaughtered cows, including 30 of non-pregnancy, and used enzyme-linked immunosorbent assay (ELISA) for estimation of plasma progesterone concentration and serum luteal hormone. The mean diameter of non-pregnant corpus luteum is $18.9{\pm}4.2{\times}15.6{\pm}3.6 mm$ and that of pregnant corpus luteum is $22.5{\pm}2.7{\times}18.7{\pm}2.9 mm$. This indicates that corpus luteum is more developed in the ovary of pregnant than non-pregnant cows (P<0.05). The diameter of pregnant corpus luteum according to the stage of pregnancy showed $21.3{\pm}2.4{\pm}18.4{\pm}2.6 mm$ in early stage (1-3 month), $23.4{\pm}2.8{\times}19.1{\pm}2.7 mm$ in middle stage (4-6 month) and $22.8{\pm}3.0{\times}18.8{\pm}2.4mm$, in last stage (7-9 month). This indicates that corpus luteum in middle and last stage is more significantly developed than that of early stage(P<0.05). The mean plasma progesterone concentration of cows showing size of non-pregnant corpus luteum was $4.58{\pm}0.92ng/ml$ and that of pregnant corpus luteum $8.26{\pm}0.98ng/ml$. Thus, it was more significantly increased in pregnant corpus luteum(P<0.02).. However, it was low to $0.58{\pm}0.39ng/ml$. in estrus (corpus albicans). The plasma progesterone concentration according to gestation period was high in proportion to the degree of development in corpus luteum and more significantly increased (P<0.05) and maintained in middle and last state than early state. The concentration was sharply decreased to $0.56{\pm}0.32ng/ml$ at parturition. As a consequence, we can practice the early pregnancy diagnosis by confirming non-pregnancy when the mean plasma progesterone concentration is below 1ng/ml 19 to 22 days after AI and this can be available to diagnose reproductive disorder.
The study was conducted on 20 Holstein X Sahiwal cross bred dairy cows, with an average milk production of $2,752{\pm}113.79$ liters in $284{\pm}5.75$ days during a single lactation, that were divided in to two groups of 10 animals. We investigated the oxidative stress and antioxidant status during the transition period in dairy cows. In this study, plasma level of MDA was considered as an indicator of lipid peroxidation and SOD, catalase, GSH and GSHPx as antioxidants. The lipid peroxidation was significantly (p<0.001) higher in cows during early lactation as compared to the cows in advanced pregnancy. A significant positive correlation (r = +0.831, p<0.01) was determined between MDA and catalase in early lactating cows. In early lactating cows, blood glutathione was significantly lower than in advanced pregnant cows. However, early lactating cows showed non-significant negative correlation for all antioxidant enzymes with lipid peroxidation. In conclusion, dairy cows seemed to have more oxidative stress and low antioxidant defense during early lactation or just after parturition than advanced pregnant cows, and this appears to be the reason for their increased susceptibility to production diseases (e.g. mastitis, metritis, retention of fetal membranes etc.) and other health problems.
The study was carried out to find out the changes of hormone levels in blood serum and milk of Holstein cows during the estrous cycle. The progesterone, estradiol-$17{\beta}$ from the blood serum and milk samples were assayed by radioimmunoassay methods. The results of this study were summarized as follows; 1. The progesterone levels in blood serum during the estrous cycles began to decline rapidly at 2 days before estrus, decreased to $0.27{\pm}0.18ng/ml$ at on the day of estrus, and reached a peak mean level of $3.33{\pm}0.47ng/ml$ at 15 days after estrus. 2. The progesterone levels in milk during the estrous cycles began to decline rapidly at 2 days before estrus, decreased to $0.80{\pm}0.18ng/ml$ on the day of estrus, and increased a peak mean level of $3.80{\pm}0.36ng/ml$ at 15 days after estrus. 3. The estradiol-$17{\beta}$ levels in blood serum during the estrous cycles showed a peak mean level of $9.79{\pm}1.72pg/ml$ on the day of estrus, and decreased from $4.79{\pm}1.82pg/ml$ to $5.73{\pm}0.96pg/ml$ at luteal phase. 4. The estradiol-$17{\beta}$ levels in milk during the estrous cycles showed a peak mean level of $36.80{\pm}2.04pg/ml$ on the day of estrus, and decreased from $18.93{\pm}0.84pg/ml$ to $19.50{\pm}1.12pg/ml$ at luteal phase. 5. During 20 to 25 days after artificial insemination, the accuracy of pregnancy diagnosis from the blood serum progesterone levels were 87.5% for non pregnant cows (<2.0ng/ml), and 83.3% for pregnant cows ($${\geq_-}$$3.0 ng/ml). The accuracy of pregnancy diagnosis from the milk progesterone levels were 75.0% for non-pregnant cows (<2.4 ng/ml), and 94.4% for pregnant cows ($${\geq_-}$$3.2 ng/ml).
The 3211 Korean native cows raised in Muan, Milyang were investigated for the purpose of finding reproductive rate of cows that retained fetal membrane for two years from April, 1981 to March, 1983. The results obtained were summarized as follows ; 1. The incidence of retained fetal membrane was found in 64 cows(1.98%). 2. The day to first estrus was similar to normal postpartum period unless concurrent genital infection occured. 3. Conception rate of cows that retained fetal membrane was 32.7%, 10.9%, 5.2% and 3.6% at the first, second, third and forth service, respectively and non pregnant cow was 47.6%. 4. Reproductive disorder of non pregnant cows was the highest in repeat breeder and the effect of treatment was very low.
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