• Title/Summary/Keyword: Estrus serum

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Pre-validation of the OECD Enhanced Test Guideline 407 Protocol on Screening and Testing for Endocrine Disrupters (Propylthiouracil을 이용한 OECD enhanced TG407의 내분비계 장애 물질검색을 위한 유효화 실험)

  • 강경선;김대용;제정환;김태원;김형섭;박지은;윤준원;김경배;이지해
    • Toxicological Research
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    • v.17 no.3
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    • pp.203-213
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    • 2001
  • We investigated the toxic effects of propylthiouracil (PTU) In Sprague-Dawley (SD) rats to develop and validate an enhanced Protocol for Test Guideline 407 as OECD Project. Twenty male and female SD rats,7 weeks old, were treated with PTU in corn oil at levels of 0, 0.1, 1 and 10 mg/kg/day for 4 weeks orally. Clinical observation, body weight changes, food uptake, water consumption, urinalysis, estrus cycle and sperm analysis, serum chemist교, autopsy findings and histopathological findings were evaluated in this study. No clinical signs and mortality were observed in the study. The body weights and food uptakes in the group treated with 10 mg/kg/day were reduced from 3 weeks after the initiation of the treatment. The levels of 3,5,3'-triiodothyronine (T3) and thyroxine (T4, 3,5,3',5'-tetraiodothyrosine) were also significantly decreased in the group treated with 10 mg/kg/day. Also, the relative and absolute organ weights of thymuses were decreased. Thyroid glands of rats in the group treated with PTU 10 mg/kg/day were bigger than those of rats in the control group. In the histopathological examination, diffuse hyperplasia and hypertrophy of thyroid follicular cells were observed in all treatment groups, leading to the reduction of lumen size and papillary enfolding of lining epithelium. The degree of lesion was increased in a dose-dependent manner. The results suggested that PTU would cause toxicity in thyroid gland and decrease the levels of T3 and T4 in SD rats. However there were no effects on the other organ including testis and uterus especially in spermatogenesis and estrus cycle. On the basis of the results, enhanced protocol for Test Guideline (TG) 407 may be sensitive and reliable to detect endocrine-active substances like PTU.

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Effect of green tea extract in extender of Simmental bull semen on pregnancy rate of recipients

  • Susilowati, Suherni;Sardjito, Trilas;Mustofa, Imam;Widodo, Oky Setio;Kurnijasanti, Rochmah
    • Animal Bioscience
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    • v.34 no.2
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    • pp.198-204
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    • 2021
  • Objective: The aim of this study was to ascertain the effects of adding green tea extract (GTE) to skim milk-egg yolk (SM-EY) extender on both the quality of post-thawed bull semen and the pregnancy rates of the recipient cows. Methods: Twelve ejaculates from four Simmental bulls, aged 3 to 5 years and weighing 900 to 950 kg, were diluted SM-EY extender, added with 0, 0.05, 0.1, and 0.15 mg GTE/100 mL extender and then frozen. After four weeks storage in liquid nitrogen, the sperm were thawed and evaluated for viability, motility, intact plasma membrane (IPM), and DNA fragmentation. Meanwhile, the estrus cycles of 48 recipient cows were synchronized by intramuscular administration of a single injection of 5 mg prostaglandin F2α. Estrus cows were divided into four equal groups and inseminated artificially 18 to 20 h after the onset of estrus by using semen from each extender group. Pregnancy was diagnosed by measuring serum progesterone levels at 21 days, followed by transrectal palpation 90 days after insemination. Results: The findings revealed that adding 0.1 mg of GTE/100 mL extender produced the highest percentages of sperm viability (70.67%±1.75%), motility (69.17%±1.47%), and IPM (69.23%±1.21%) and the lowest percentage of DNA fragmentation (3.00%±0.50%). The pregnancy diagnosis revealed that all cows (36/36) inseminated using frozen semen in GTE addition extender were pregnant (pregnancy rate 100%), whereas the pregnancy rate of the control group was 83.33% (10/12). Conclusion: It may be concluded that 0.1 mg GTE/100 mL extender yields the best quality of spermatozoa and that all variants doses of GTE in extender produce a higher pregnancy rate among recipient cows.

Development a Following of Serum Addition In Vitro Culture and Embryo Transfer (체외 배양액의 조성과 혈청의 첨가가 한우 체외 수정란의 발달과 임신율에 미치는 효과)

  • Choi, Soo-Ho;Park, Yong-Soo;Lee, Joon-Hee;Kang, Tae-Yeong;Kim, Joo-Heon;Rho, Gyu-Jin
    • Journal of Embryo Transfer
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    • v.23 no.1
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    • pp.51-57
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    • 2008
  • The present study investigated the efficient methods to produce in vitro Hanwoo embryos, and to improve the pregnancy rate. The developmental rate, total cell number and ICM ratio of in vitro embryos were compared amongst different culture media. Comparisons were also made on the status of recipients, pregnancy rate along with day of transfer after the estrus. Development of embryos into blastocyst stage in IVMD101 supplemented with 5% fetal bovine serum (FBS) group was significantly higher (34.2%) than that of TCM-199 supplemented with 5% FBS (26.8%) and IVMD101 without FBS (25.9%) (p<0.05). The development rate to blastocyst stage was significantly faster in IVMD101(5% FBS) than that of other groups ($0.2{\sim}2.3%$) (p<0.05). The average number of inner cell mass and trophectoderm were similar among treatment groups, which were $36.0{\sim}44.7$ and $83.3{\sim}106.7$. However, total cell number in IVMD101(5% FBS and 0% FBS) was significantly higher than that of TCM199(5% FBS). There were no differences in the pregnancy rate among treatment groups (32.0%, 33.9% and 28.6%, respectively). However, the pregnancy rate of Day 6 embryos cultured in IVMD101(5% FBS) was significantly (p<0.01) higher than IVMD101 without FBS and TCM-199 + 5% FBS (38.0% vs. 17.2% and 32.4%, respectively). No significant difference was observed for the pregnancy rate between heifer and cow transferred with Day 6 embryos cultured in IVMD 101(5% FBS) (42.7% and 39.3%, respectively). However, there was a significant difference of pregnancy rate (p<0.05) in heifer between one and two embryos transferred (31.4% and 41.9%). There was no difference of pregnancy rate among transfer days after estrus between heifer and cow, but the pregnancy rate of transfer to heifer with day 6 after estrus was significantly higher (p<0.05) than that of day 7 and 8 (22.2% vs. 49.0% and 38.7% respectively). Based on the above findings, there is a possibility to produce in vitro produced embryos cultured in IVMD101(5% FBS) showed higher blastocyst rate and the increased cell number. In terms of the pregnancy rate of in vitro produced embryos, the highest pregnancy rate was observed when two embryos were cultured in IVMD101(5% FBS) and transferred.

Alteration in Concentrations of Leptin and Insulin in Korean Cattle with Reproductive Disorders (번식장애 한우의 렙틴 및 인슐린 농도 변화)

  • Choe, Chang-Yong;Kang, Da-Won
    • Journal of Embryo Transfer
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    • v.23 no.3
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    • pp.189-195
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    • 2008
  • The objective of this study is to investigate the changes in concentrations of leptin and insulin in serum of Korean cattle (Hanwoo) with reproductive disorders and to examine the relationship among leptin, insulin, and body condition score (BCS). The concentration of leptin in serum of pregnant Hanwoo showed insignificant difference from that in serum of Hanwoo with reproductive disorder, such as repeat breeding, follicular cyst, corpus luteum cyst, ovarian atrophy, and feeble estrus (p>0.05). However, the concentrations of leptin and insulin in serum were changed with different BCS value. In emaciated Hanwoo (BCS $2.0\sim2.9$), they were significantly decreased compared to BCS $3.0\sim3.4$ (p<0.05). The leptin showed different genotypes with different BCS value. In BCS $2.0\sim2.9$, C/T genotype was expressed (83.3%) more than C/C (16.7%) or T/T (0%) genotype, whereas C/C genotype was expressed (62.5%) more than C/T (25.0%) or T/T (12.5%) genotype in BCS $3.5\sim4.0$. The insulin concentration in follicular fluid obtained from ovary with follicular cyst which has follicles having diameter of $25\sim40 mm$ was significantly higher (p<0.05) than those in normal follicle fluid which has follicles having diameter of $3\sim10 mm$. These results showed that concentration of leptin and insulin in serum were related to BCS value and follicular size and suggest that the changes in concentration of leptin and/or insulin in serum could be a potent biomarker for diagnosis of bovine reproductive disorder.

Enzyme-linked immunosorbent assay(ELISA) for determination of progesterone concentration in bovine serum (ELISA에 의한 소의 혈청 progesterone 농도 측정)

  • Kang, Chung-boo;Son, Min-soo;Lee, Eun-sug;Cho, Kyu-woan;Kim, Chur-ho
    • Korean Journal of Veterinary Research
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    • v.33 no.3
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    • pp.539-545
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    • 1993
  • This study was carried out the determine the progesterone concentration for serum by enzyme-linked immunosorbent assay(ELISA) in bovine adult at estrous, pregnant, after patuation and male, female calves of 1 month old, respectively. The results obtained were summarized as follows ; 1. The assay has a sesitivity of $0.1ng/m{\ell}$. 2. Intra and inter-assay coefficient of variation were 4.5%, 6.1~9.4% when used for the determination of progesterone in samples of bovine serum. 3. The percentages of recovery for progesterone added were between 88.0 to 88.9%. 4. Progesterone concentration of adult bovine serum at estrus, pregnant and after 1 day of parturition were $0.37{\pm}0.16$, $7.1{\pm}1.0$, $0.13{\pm}0.02ng/m{\ell}$, respectively. 5. There was no differences in serum progesterone concentration of calves both male($0.16{\pm}0.03ng/m{\ell}$) and female($0.15{\pm}0.04ng/m{\ell}$) on 1 month old. From these results, progesterone determination by ELISA is very applicable to detect of early pregnancy diagnosis, factorial analysis of reproductive disorder, and also reproductive physiological functions such as veterinary therapeutic measures and monitoring of cyclicity.

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In Vitro Nuclear Maturation of Canine Oocytes Obtained from Differents Stage of Estrus Cycle (개의 발정주기가 난자의 체외성숙에 미치는 영향)

  • Kim, Min-Kyu;Kim, Hye-Jin;Cho, Jong-Ki;Jang, Gu;Lee, Kyu-Seung;Kang, Sung-Geun;Lee, Byung-Chun;Hwang, Woo-Seok
    • Journal of Embryo Transfer
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    • v.17 no.2
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    • pp.145-151
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    • 2002
  • The aim of these experiments was to investigate in vitro nuclear maturation of canine oocyte collected from various stages of estrus cycles, Ovaries were obtained from 1 to 4 year-old mongrel bitch and minced for oocyte collection in phosphate buffered saline with 100 iu penicillin-G $m\ell$/sup -1/, 50 $\mu\textrm{g}$ streptomycin sulphate $m\ell$/sup -1/ and 0.1% (w/v) polyvinyl alcohol. Cumulus-oocyte-complexes (COCs) were washed in HEPES buffered tissue culture medium (TCM)199 and in vitro matured in TCM-199 culture medium supplemented with sodium pyruvate 0.028mg/$m\ell$, L-glutamine 0.146mg/$m\ell$, penicillin G 10,000 IU/$m\ell$, streptomycin 0.031mg/$m\ell$ and 10% (v/v) fatal calf serum. COCs were in vitro matured for 48~72 hrs at 39$^{\circ}C$ in humidified 5% $CO_2$ in air atmosphere. In vitro matured oocytes were remove the cumulus cells using 0.2% (v/v) hyaluronidase. After denuding, oocyte were placed in acetic acid : methanol : chlorform solusion (3:6 : 1.5 v/v) for 30 sec and acetic acid: ethanol(1:3 v/v) for 48hrs fixation. Nuclear maturation was classified to GV, GVBD, MI, MII and degenerate oocyte under microscopy after 1% aceto-orcein stain. In vitro maturation rates at 48hrs were not significantly difference among the oocytes collected from different stage of estrus at 15.9%, 16.3%, 23.7% and 18.2% for anestrus, proestrus, estrus and diestrus. However, the oocytes maturation(36.6%) of collected from estrus ovaries were significantly different from oocytes derived from proesturs, diestrus and anestrus ovaries(30.8%, 17.5% and 22.1%; p<0.05). The overall in vitro maturation rates were significantly higher (p<0.05) in 72hrs culture than 48hrs culture system. In summary, there was a tendency for higher in vitro maturation rates with the oocyte collected from estrus ovary than other stages of estrus. Also, for nuclear maturation, in vitro culture of oocyte for 72hrs was better than 48hrs culture.

Association of Endocrine Factors (Insulin-Like Growth Factor-II and Binding Protein-3) with Litter Size in Pigs

  • Yun, J.S.;Kang, W.J.;Seo, D.S.;Park, S.S.;Hong, K.C.;Lee, C.Y.;Ko, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.3
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    • pp.307-315
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    • 2001
  • Litter size has been one of the important economic traits in porcine reproduction. The insulin-like growth factor (IGF) system has been shown to mediate actions of the steroid hormone or to synergize with other endocrine factors so that it consequently plays roles in reproductive processes, including ovulation, implantation, maintenance of pregnancy, and fetal development. However, the effect of the serum IGF system on porcine litter size has not been deeply studied. Therefore, this study was conducted to relate serum IFG-II concentration and IGF binding protein-3 (IGFBP-3) expression with porcine litter size. Moreover, the possible association of those with estrogen receptor (ER) as a candidate gene for litter size was investigated. Swine were separated into two groups showing high and low litter sizes, and sera were collected from sows in the estrous cycle to postnatal growth of their female progeny. Serum IFG-II concentration was measured by radioimmunoassay and IGFBP-3 expression was detected by Western ligand blotting. During the estrous cycle, IGFBP-3 expression in both groups decreased moderately from metestrus to estrus, but IFG-II concentration showed a reverse pattern. Also, IFG-II concentration and IGFBP-3 expression decreased gradually as pregnancy proceeded. Unlike IGFBP-3, IFG-II decreased moderately as newborn pigs grew. Significant differences in serum IFG-II amount between the two groups were detected at 60 (p<0.01), 75, 90, and 105 d (p<0.05) of pregnancy and at 60 (p<0.01), 45, and 105 d (p<0.05) of postnatal growth. Furthermore, based on ER genotypes, a high litter size group with genotypes AB and BB showed lower IFG-II concentration than a low litter size group with a genotype AA during pregnancy. Taken together, the results indicate that the serum IFG-II and IGFBP-3 are correlated with the litter size in pigs.

The Estrous Cycle of the Markhoz Goat in Iran

  • Farshad, A.;Akhondzadeh, M.J.;Zamiri, M.J.;Sadeghi, GH.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.10
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    • pp.1411-1415
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    • 2008
  • The purpose of the present study was to determine the onset of breeding season, the occurrence of silent and true heats and the duration of estrus in female Markhoz goats of the Kurdistan province in Iran. Ten, 3 years-old Markhoz does with an average weight of $34.05{\pm}2.62kg$ and with one kidding record, were used. The goats were maintained in an open barn under constant nutritional levels and natural photoperiod. One aproned buck was used twice a day every 12 h to detect estrus from mid August to early January. For the determination of the onset of reproductive activity as well as occurrence of silent heat, blood samples were collected every 10 days, from the beginning of the experiment. After $2^{nd}$ standing heat, blood samples were obtained twice a week in order to assess luteal activity and the length of estrous cycles. In this study, estrous behavior was observed including sniffing, vocal exchange, following courtship, flehman, standing heat and mounting. The results of the progesterone assay indicated that in this goat silent heats occur mostly in the early breeding season. The first standing heat was observed in mid-October which was considered as the onset of the breeding season. Duration of the estrous cycle and estrus was recorded as being $20.93{\pm}1.56days$ and $38.86{\pm}15.19h$, respectively. The correlation coefficient between length of daylight and occurrence of estrus was negative (r = -0.470) but not significant (p>0.05). The data showed that there was no significant effect of body weight on estrous cycles (first, second and third) and estrous periods (first, second, third and fourth). Progesterone levels were not significantly different in the first, second and third estrous cycles at days 0, 4, 10, 12, 14 and 19. The results of progesterone assay during the estrous cycle indicate that follicular and luteal phases last 4-5 and 14-15 days, respectively and the concentration of serum progesterone in these phases was $0.88{\pm}0.08$ and $7.44{\pm}0.26ng/ml$, respectively. The study concluded that Markhoz does could be considered as a breed with a short breeding season and an optimal estrous activity in autumn.

Estrus Induction Rates and Conception Rates by MGA Feeding System in Hanwoo (MGA Feeding System에 의한 발정 동기화 방법이 한우의 발정 발현율과 수태율에 미치는 영향)

  • Lee, M.S.;Jung, Y.H.;Park, S.B.;Im, S.K.;Park, J.J.;Jung, J.K.;Moon, S.J.;Kim, C.K.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.177-182
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    • 2006
  • This study was conducted to investigate the effects of melengesterol acetate (MGA) and $PGF_{2{\alpha}}$ administrations on serum progesterone level, synchrony of estrus and conception rates in Han-woo. Firstly, ten heifers and one freematin were fed 0.5 mg MGA/day for 14 days in a grain carrier, and after 19 days of MGA feeding, a single injection of 25 mg $PGF_{2{\alpha}}$ were treated. Blood samples were collected to evaluate serum progesterone concentrations from the start of feeding of MGA until the end of feeding and subsequent estrous detection and artificial insemination (AI) at 3 days intervals, and on days of $PGF_{2{\alpha}}$ injection, estrous detection, AI, and 15th and 60th days after AI. The level of progesterone in the blood began to increase from 7 days after MGA feeding, and 9 days after feeding it became 5.4 ng/ml and maintained that level thereafter. On the 33th day when the $PGF_{2{\alpha}}$ was injected, it reached the peak level of 7.6 ng/ml. However, 2-3 days after $PGF_{2{\alpha}}$ injection, it dropped to 1.4 ng/ml drastically (p<0.05). Secondly, one hundred and ninety four Hanwoo heifers or cows were divided into two groups to compare estrous induction and conception rates: the one treated with MGA and $PGF_{2{\alpha}}$, (n=104) and the other with $PGF_{2{\alpha}}$ treatment (two injections at 11 days interval, n=90). The heifers or cows treated with MGA and $PGF_{2{\alpha}}$ were identical to those used as above. The percentages of heifers or cows showed estrus were higher in the $MGA+PGF_{2{\alpha}}$ treatment (91.3%) than in the $PGF_{2{\alpha}}$ treatment (72.2%, p<0.05). Conception rates were also higher in the $MGA+PGF_{2{\alpha}}$ treatment (94.2%) than in the $PGF_{2{\alpha}}$ treatment (88.9%, p<0.05). The results of this experiment indicate that estrus synchronization using $MGA+PGF_{2{\alpha}}$ is more effective than that using $PGF_{2{\alpha}}$ (two injections) in Hanwoo.

Effect of Serum IGF-I on Progesterone Concentration during Early Pregnancy in Korean Native Cattle (Hanwoo)

  • Ryu, Yang-Hwan;Yang, Jong-Yong;Seo, Dong-Sam;Ko, Yong
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.2
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    • pp.176-179
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    • 2003
  • Insulin-like growth factor-I (IGF-I) is a polypeptide that has the function of regulating the expression of steroid hormones through endocrine, paracrine, and autocrine actions in reproductive organs. Moreover, IGF-I is involved in ovulation, implantation, maintenance of pregnancy, and development of fetuses in animals. Therefore, this study was conducted to investigate the effects of serum IGF-I concentration on progesterone ($P_4$) concentration and pregnancy rates in Korean native cattle (Hanwoo). Blood was collected at estrus (Day 0) and Day 11. Artificial insemination was performed at Day 0. Serum IGF-I and $P_4$ concentrations were measured by radioimmunoassay (RIA). Overall, $P_4$ concentration was higher at Day 11 than Day 0, whereas the pattern of IGF-I concentration was reversed. When animals were divided into two groups depending on the pregnancy status, $P_4$ concentrations of the pregnant group was significantly higher than that of the non-pregnant group at Day 0 (p<0.05) and Day 11 (p<0.05). But, lower IGF-I concentrations were detected in the pregnant group at Day 0 (p<0.05) and Day 11 (p<0.05) compared to the non-pregnant group. In conclusion, these results indicated that serum IGF-I is inversely associated with $P_4$ concentration during early pregnancy in Hanwoo.