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In vitro Fertilization and Development of Pig Oocytes Inseminated with Boar Sperm by Different Sperm Washing Media after Thawing of the Frozen Straws

  • Yi, Y.J.;Ko, H.J.;Lee, S.H.;Yang, C.B.;Son, D.S.;Kim, H.K.;Park, C.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.2
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    • pp.164-167
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    • 2004
  • This study was carried out to investigate in vitro fertilization and development of in vitro matured pig oocytes inseminated with the Duroc boar sperm by different sperm washing media after thawing of the 5 ml frozen straws. Immature follicular oocytes (30-40) were transferred into each well of a Nunc 4-well multidish containing $500{\mu}l$ mTCM199 maturation medium. The sperm rich portion of ejaculates was collected into a 250 ml insulated vacuum bottle and gradually cooled 22 to $24^{\circ}C$ over a 2 h period. Semen was centrifuged at 800 g for 10 min and the seminal plasma discarded. Sperm were esuspended in a lactose-egg yolk and N-acetyl-Dglucosamine (LEN) diluent to contain $1{\times}10^{9}$ sperm/ml and cooled to $5^{\circ}C$ over a 2 h period. Immediately before freezing, semen was rediluted with an equal volume of LEN+4% glycerol and packed into 5 ml straws. After thawing of the 5 ml straw, the 5 ml semen was diluted with 20 ml Beltsville thawing solution (BTS) at room temperature. Oocytes were inseminated with untreated (unwashed and nonpreincubated) or treated sperm (washed two times in BTS, mTLP-PVA and mTBM media, respectively and nonpreincubated) with $2{\times}10^{7}$ sperm concentration. Oocytes were coincubated for 6 h in $500{\mu}l$ mTBM fertilization. At 6 h after IVF, oocytes were transferred into $500{\mu}l$ NCSU-23 culture medium for further culture of 6 h. Sperm penetration, polyspermy and male pronuclear formation of oocytes at 12 h after IVF and developmental ability of oocytes at 48 h after IVF were evaluated. Sperm penetration rate, male pronuclear formation and rate of cleaved embryos were higher in the BTS, mTLP-PVA and mTBM treatments than the unwashed treatment (p<0.05). The rate of blastocysts from the cleaved oocytes (2-4 cell stage) were higher in the mTLP-PVA treatment than in the unwashed, BTS and mTBM treatments. In conclusion, we recommend the washing of frozen-thawed sperm with mTLP-PVA medium before in vitro fertilization of oocytes in mTBM medium.

The Effects of Chinese and Argentine Soybeans on Nutrient Digestibility and Organ Morphology in Landrace and Chinese Min Pigs

  • Qin, G.X.;Xu, L.M.;Jiang, H.L.;van der Poel, A.F.B.;Bosch, M.W.;Verstegen, M.W.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.4
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    • pp.555-564
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    • 2002
  • Twenty Landrace and twenty Min piglets, with an average initial body weight of 22.4 kg, were randomly divided into 5 groups with 4 animals per group, within each of the breeds. The piglets were housed in individual concrete pens. Each group of the piglets was fed one of 5 diets. The diets contained either 20% raw Argentine soybeans, 20% processed Argentine soybeans ($118^{\circ}C$ for 7.5 min.), 20% raw Chinese soybeans, 20% processed Chinese soybeans ($118^{\circ}C$ for 7.5 min.) or no soybean products (control diet). Faecal samples were collected on days 6, 7 and 8 of the treatment period. Digestibilities of dietary nutrients were determined with AIA (acid insoluble ash) as a marker. After a 17 day treatment, three piglets were killed from each of the groups. Tissue samples of small and large intestine for light and electron microscopy examination were taken immediately after the opening of abdomen. Then, the weight or size of relevant organs was measured. The results show that the digestibilities of dry matter (DM), crude protein (CP) and fat were higher in Min piglets than in Landrace piglets (p<0.05). The diets containing processed soybeans had a significant higher CP digestibility than the control diet and the diets containing raw soybeans (p<0.05). Landrace piglets had heavier and longer small intestines, heavier kidneys and a lighter spleen than Min piglets (p<0.05). The pancreas of the animals fed the diets containing processed soybeans was heavier than that of the animals fed control diet (p<0.05) and the diets containing raw soybeans. But, the differences between raw and processed soybean diets were not significant. A significant interaction (p<0.05) between diet and pig breed was observed in weight of the small intestine. The Landrace piglets increased the weight in their small intestine when they were fed the diets containing soybeans. In the light micrographs and electron scanning micrographs, it was found that the villi of small intestinal epithelium of animals (especially Landrace piglets) fed the diets containing raw Chinese soybeans were seriously damaged. The transmission electron micrograph showed that a lot of vesicles were located between the small intestinal microvilli of these piglets. The histological examination also indicated that the proportion of goblet cells in villi and crypts in the piglets consuming the control diet was significantly lower (p<0.01 and p<0.02, respectively) than those of the animals consuming the diets containing raw or processed soybeans.

Retrospective Analysis of Neoadjuvant Chemotherapy for Breast Cancer in Turkish Patients

  • Duman, Berna Bozkurt;Afsar, Cigdem Usul;Gunaldi, Meral;Sahin, Berksoy;Kara, I. Oguz;Erkisi, Melek;Ercolak, Vehbi
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4119-4123
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    • 2012
  • Background: Neoadjuvant systemic chemotherapy is the accepted approach for women with locally advanced breast cancer. Anthracycline- and taxane-based regimens have been extensively studied in clinical trials and consequently are widely used. In this study aimed to research the complete response (pCR) rates in different regimens for neoadjuvant setting and determine associated clinical and biological factors. Methods: This study included 63 patients diagnosed with breast carcinoma among 95 patients that had been treated with neoadjuvant chemotherapy between 2007 and 2010. TNM staging system was used for staging. The histologic response to neoadjuvant chemotherapy was characterized as a pCR when there was no evidence of residual invasive tumor in the breast or axillary lymph nodes. Biologic subclassification using estrogen receptor (ER), progesterone receptor (PR), HER2 were performed. Luminal A was defined as ER+, PR+, HER2-; Luminal B tumor was defined as ER+, PR-, HER2-; ER+, PR-, HER2+; ER-, PR+, HER2-; ER+, PR+, HER2+; HER2 like tumor ER-, PR+, HER2+; and triple negative tumor ER, PR, HER2 negative. Results: Patients median age was 54.14 (min-max: 30-75). Thirty-two patients (50.8%) were premenapousal and 31 (49.2%) were postmenapousal. Staging was performed postoperatively based on the pathology report and appropriated imaging modalities The TNM (tumor, lymph node, metastasis) system was used for clinical and pathological staging. Fifty-seven (90.5%) were invasive ductal carcinomas, 6 (9.5%) were other subtypes. Thirty nine (61.9%) were grade II and 24 (38.1%) were grade III. Seven (11.1%) patients were stage II and 56 (88.9) patients were stage III. The patients were classified for ER, PR receptor and HER2 positivity. Seventeen patients had complete response to chemotherapy. Forty patients (63.5%) were treated with dose dense regimen (cyclophosphamide 600 mg/m2 and doxorubicine 60 mg/m every two weeks than paclitaxel 175 mg/m2 every two weeks with filgrastim support) 40 patients (48%) were treated anthracycline and taxane containing regimens. Thirteen patients (76%) from 17 patients with pCR were treated with the dose dense regimen but without statistical significance (p=0.06). pCR was higher in HER2(-), ER(-), grade III, premenopausal patients. Conclusion: pCR rate was higher in the group that treated with dose dense regimen, which should thus be the selected regimen in neoadjuvant setting. Some other factors can predict pCR in Turkish patients, like grade, menopausal status, triple negativity, percentage of ER positivity, and HER2 expression.

Plant Diversity, Tree Regeneration, Biomass Production and Carbon Storage in Different Oak Forests on Ridge Tops of Garhwal Himalaya

  • Sharma, Chandra Mohan;Tiwari, Om Prakash;Rana, Yashwant Singh;Krishan, Ram;Mishra, Ashish Kumar
    • Journal of Forest and Environmental Science
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    • v.32 no.4
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    • pp.329-343
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    • 2016
  • The present study was conducted on ridge tops of moist temperate Oak forests in Garhwal Himalaya to assess the plant diversity, regeneration, biomass production and carbon assimilation in different Oak forests. For this purpose, three Oak forest types viz., (a) Quercus leucotrichophora or Banj Oak (FT1; between 1,428-2,578 m asl), (b) Quercus floribunda or Moru Oak (FT2; between 2,430-2,697 m asl) and (c) Quercus semecarpifolia or Kharsu Oak (FT3; between 2,418-3,540 m asl) were selected on different ridge tops in Bhagirathi catchment area of Garhwal Himalaya. A total of 91 plant species including 23 trees (8 gymnosperms and 15 angiosperms), 21 shrubs and 47 herbs species belonging to 46 families were recorded from all the ridge top Oak forests. The highest mean tree density ($607{\pm}33.60trees\;ha^{-1}$) was observed in Q. floribunda forest with lower mean total basal cover (TBC) value ($48.02{\pm}3.67m^2ha^{-1}$), whereas highest TBC value ($80.16{\pm}3.30m^2ha^{-1}$) was recorded for Q. semecarpifolia forest, with lowest mean stem density ($594{\pm}23.43stems\;ha^{-1}$). The total biomass density (TBD) across three Oak forests ranged between $497.32{\pm}83.70$ (FT1) and $663.16{\pm}93.85t\;ha^{-1}$ (FT3), while the total carbon density (TCD) values ranged between $228.75{\pm}22.27$ (FT1) and $304.31{\pm}18.12t\;ha^{-1}$ (FT3). Most of the tree species were found with good regeneration (GR) status (average 45%) in all the forest types whereas, few species were found not regenerating (NR) (average 17%) however, few new recruitments were also recorded. ANOVA (Post-Hoc Tukey's test at 5% significance level) indicated significant forest-wise differences in TBC, TBD and TCD (in tree layer); family and evenness (in shrub layer only) values, while insignificant differences were noticed in density values of tree, seedling and herb layer.

The Effects of Plum Extracts on the Proliferation of Human Epithelial Cell and Human Cervical Cancer Cells (자두 추출물이 인체 상피세포와 자궁경부암세포의 증식에 미치는 효과)

  • Han, Man-Deuk;Kweon, Dur-Han;Kang, Byung-Tae;Lee, Jae-Woo;Yoon, Ok-Hyun
    • Journal of the East Asian Society of Dietary Life
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    • v.17 no.5
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    • pp.710-718
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    • 2007
  • This study was designed to investigate the effects of plum(Prunus salicina Lindl. cultivars 'Oishiwase', 'Formosa', and 'Soldam') extracts on the proliferation as well as inhibition of human epithelial cells(HaCaT), human cervical carcinoma (HeLa, SiHa, and C33A) cells, and human stomach adenocarcinoma(SNU 638) cells. Dried plum was sequentially extracted and fractionated by hexane(KC-01), chloroform(KC-02), ethyl acetate(KC-03), n-butanol(KC-04), water(KC-05), methanol(KC-6), and hot water extract(KC-07). The epithelial and cancer cells were exposed for 48 h to $50{\mu}g/mL$ of plum extract in vitro, and were then analysed by a sulforhodamin B(SRB) staining assay. The methanol extract(KCP-6) of 'Formosa' proliferated not only the HaCaT cells(147.3%), but also the cervical carcinoma C33A cells(167.8%). The ethyl acetate extract of 'Soldam'(KCJ-3) significantly reduced the proliferation rate of the HPV positive conical carcinoma cells, at 61.5% for the SiHa cells and 70.5% for the HeLa cells. In the C33A cells, which are HPV negative cervical carcinoma cells, the hexane fractions of 'Formosa'(KCP-1) and 'Oishiwase'(KCD-1) markedly suppressed proliferation activity at 20.4% and 61.7%, respectively. However, the proliferation rate of the normal epithelial cells(HaCaT cell) was not reduced the proliferation rate by KCJ-3, KCP-1, or KCD-1, There were no significant effects on proliferation of the stomach cancer cells(SNU 638) by any of the extracts or fractions of the plum cultivars. These results suggest that the anti-proliferative effects of the plum cultivars were selective to the cancer cell origin. In conclusion, we found that several plum cultivar extracts, especially, the ethyl acetate fraction of 'Soldam" and the hexane fraction of "Formosa', have anti-proliferative activity toward human cervical carcinoma cells. However, further investigation is needed to assess the molecular mechanisms that mediate the antiproliferation activities of the plum cultivars.

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Seismic Response Control of a Cable-Stayed Bridge Using Passive, Active, Semiactive and Hybrid Systems (수동, 능동, 반능동 및 복합 시스템을 이용한 사장교의 지진응답 제어)

  • ;;Spencer, B. F.
    • Journal of the Earthquake Engineering Society of Korea
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    • v.7 no.1
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    • pp.17-29
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    • 2003
  • This paper preliminarily investigates the effectiveness of various control systems, such as passive, active, semiactive and hybrid control, for seismic protection of cable-stayed bridges by examining the ASCE first generation benchmark problem for a cable-stayed bridge. This benchm.0.00000ark problem considers the cable-stayed bridge that is scheduled for completion in Missouri, USA In 2003. Seismic considerations were strongly considered in the design of this bridge due to location of the bridge and its critical role as a principal crossing of the Mississippi River. Based on detailed drawings of this cable-stayed bridge, a three-dimensional linearized evaluation model has been developed to represent the complex behavior of the bridge. A set of eighteen evaluation criteria has been developed to evaluate the capability of each control system. In this study, four passive control systems, one active control system, two semiactive control systems and three hybrid control systems are considered. Numerical simulation results show that all the control systems are effective in reducing the responses of the benchmark cable-stayed bridge under the historical earthquakes. To get good performance, however, the passive control systems need quite large control forces compared to other control systems. The simulation results also demonstrate that the passive, semiactive and hybrid control systems are robust to the stiffness uncertainty of the structure. Therefore, the semiactive and hybrid control systems are more appropriate in real applications for full-scale civil infrastructures.

A study on dental hygiene department students' attitude toward infection in a dental hygiene workshop (치위생 실습실에서 치위생과 학생의 감염에 관한 태도 연구)

  • Kim, Eun-Hee;Kim, Seol-Ak
    • Journal of Korean society of Dental Hygiene
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    • v.9 no.4
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    • pp.849-863
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    • 2009
  • Objectives : This study was to study dental hygiene department students' management of infection and their attitude toward infection. Methods : This study was conducted from August 24, 2009 to September 20, 2009. 269 sophomore and junior college students enrolled in the department of dental hygiene from schools located in Gyeonggi-do and Gangwon-do participated in the study. A self-administered questionnaire survey was conducted. Results : 1. As for dental hygiene department students' recognition of the causes of infectious diseases, 80.7% of the respondents said hepatitis B was the infectious disease highly likely to occur in laboratories. 35.3% was correct about the antecedent variables of infection in the workshop, 24.9% about the oral symptoms of HIV and 18.6% about the diseases induced by HBV. The recognition rates were generally low. 2. About whether the respondents ask patients questions about infectious diseases, 80.7% of them answered Yes, but only 56.2% of them said they do so every time, and 17.8% of them said they never do so. 3. As far as washing hands to prevent infectious diseases is concerned, 97.4% of the respondents said hand washing helps prevent infection. 72.5% of them said they wash their hands every time before they practice on a patient, while 84.0% of them said they washed their hands after the lab practice. 90.7% said they use liquid soap containing anti-microbial agents, and 81.8% of them said they use paper towels. 4. With regard to protective gear for prevention of infectious diseases, 98.9% of the respondents said it is desirable to use disposable protective gear for each patient. When it comes to what they actually used as protective gear, 91.1% said aprons, 89.2% gloves, and 87.7% masks. However, a low percentage of the respondents actually use goggles and replace masks when they got damp, 11.2% and 24.2% respectively. Conclusions : As for treating the surface of equipments to prevent infectious diseases, most of the respondents exhibited a high recognition rate. Relative fewer respondents actually treat the surface of equipments than those respondents who are aware of the need to do so. A high percentage of the respondents also said they use alcohol sponge to treat the surface of each equipment in order to prevent infectious diseases.

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Evaluation of Greenhouse Gas Emissions from Animal Manure Treatment Systems with Life Cycle Assessment : A Case Study (전과정평가를 이용한 가축분뇨 처리시설의 온실가스 배출량 평가 : 사례 연구)

  • Park, K.H.;Choi, D.Y.;Cho, S.B.;Yang, S.H.;Hwang, O.H.
    • Journal of Animal Environmental Science
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    • v.17 no.sup
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    • pp.1-6
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    • 2011
  • Korean Government announced 'The Roadmap to realize a low carbon green society on year 2020' on July 12, 2011 in order to mitigate greenhouse gas (GHG) emissions. Non-energy category of Food, Agriculture, Forestry and Fishery (FAFF) should mitigate 1,349 kilo $CO_2$-equivalent ($CO_2$-eq.) tonnes which is 7.1% of Business-As-Usual on year 2020. The mitigation from animal manure treatment system (AMTS) comprises ca. 45% of the total mitigated amount of Non-energy category of FAFF. Hence, the precise evaluation of GHG emissions from AMTS is important to find effective mitigation measures. Life cycle assessment was used to evaluate GHG emissions from AMTS. The most GHG emitter was a composting/liquid fertilizer/activated sludge system (1,649.45 kg $CO_2$-eq./head/year) and the least GHG emitter was a activated liquid fertilizer system (1,024.46 kg $CO_2$-eq./head/year). Thermophilic oxic process showed the highest ratio (34.9%) of GHG emissions by the use of electricity to total GHG emissions from systems. Energy efficiency should be considered to mitigate GHG emissions from AMTS.

Fine Structures of the Enteroendocrine Cells in the Duodenal Mucosa of the Hedgehog, Erinaceus koreanus (고슴도치 십이지장 점막의 장내분비세포의 미세구조)

  • Choi, Wol-Bong;Won, Moo-Ho;Seo, Ji-Eun
    • Applied Microscopy
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    • v.17 no.1
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    • pp.83-97
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    • 1987
  • In order to discriminate the enteroendocrine cell types in the mucosal epithelium of the normal duodenum of the Korean hedgehog (Erinaceus koreanus). The tissues were fixed in the mixture of 1% paraformaldehyde and 1% glutaraldehyde in phosphate buffer (pH 7.2), and postfixed in 2% osmium tetroxide (phosphate buffer, pH 7.2). They were embedded in Araldite, and the ultrathin sections were made by LKB-V ultratome following the inspection of semithin sections stained with toluidine blue-borax solutions. Ultrathin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100B electron microscope. At least six types of enteroendocrine cells distributed in the mucosal epithelium of the duodenum were identified according to their morphological characteristics mainly based on the size, shape, number and electron density of the secretory granules. Type I cells had moderately developed organelles. The secretory granules were pleomorphic ($370X510nm$), and the granule cores with high electron density were enveloped in limiting membrane and characterized by a narrow halo. Type II cells contained an indented nucleus and well-developed organelles. The secretory granules were round (350 nm) and classified in two kinds by electron density, moderate and high. Both granules were surrounded by limiting membrane and those with high electron density showed often a wide halo. Type III cells had an indented nucleus. The secretory granules with various electron density were round (220 nm) in shape. The granules with high electron density were enveloped in limiting membrane and characterized by a narrow halo, but those with low or moderate electron density had not been observed the limiting membrane. Type IV cells contained an indented nucleus and moderately developed organelles. The secretory granules were round (180 nm) in shape, and the granule cores with high electron density were enveloped in limiting membrane and showed often a wide halo. Type V cells had a large amount of rough endoplasmic reticulum. Secretory granules with low or moderate electron density were round (230 nm) in shape, and surrounded by limiting membrane and showed a narrow halo. Type VI cells contained an oval nucleus and well-developed organelles, especially Golgi complex. The secretory granules with high electron density were round (210 nm) in shape. The granules were enveloped in limiting membrane and showed often a wide halo.

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Use of ultrasonography for improving reproductive efficiency in cows II. Comparative evaluation of ovarian structures using ultrasonography and plasma progesterone analysis in subestrous dairy cows (초음파 진단장치를 이용한 축우의 번식효율증진에 관한 연구 II. 무발정 젖소에서 초음파검사 및 progesterone 농도측정에 의한 난소 구조물의 비교평가)

  • Son, Chang-ho;Kang, Byong-kyu;Choi, Han-sun;Kang, Hyun-gu;Paik, In-seok;Suh, Guk-hyun
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.642-651
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    • 1998
  • The accuracy of ultrasonography for determining the presence of a functional corpus luteum in subestrous dairy cows was investigated, using a radioimmunoassay for progesterone in plasma. Luteal status (high or low progesterone concentrations) was diagnosed in 534 cows, using B-mode transrectal ultrasonography. Accuracy of ultrasonography was 96.3% and 88.8% in the cows with and without functional corpus luteum, respectively. In 362 cows diagnosed with functional corpus luteum by ultrasonographic examination, 20 cows were diagnosed with the non-functional corpus luteum by analysis of plasma progesterone concentrations (false positive). In 172 cows with non-functional corpus luteum by ultrasonographic examination, 13 cows were diagnosed with the functional corpus luteum based on plasma progesterone assay (false negative). Most of the corpus luteum with well-defined border and homogeneous echotexture were diagnosed with functional corpus luteum. All cows that were not detected a corpus luteum by ultrasonographic examination were diagnosed as non-functional corpus luteum. The corpus luteum of cows that were diagnosed with false positive appeared homogeneous echotexture and above 15 mm in diameter, but the corpus luteum was the non-functional corpus luteum within Day 5 (Day 0 is ovulation day) or after Day 19. The corpus luteum of cows that were diagnosed with false negative appeared heterogeneous echogenicity and below 15 mm in diameter, but the corpus luteum was the functional corpus luteum after Day 5 or around Day 17. It was concluded that accuracy of ultrasonography was excellent for determining the presence of a functional corpus luteum in subestrous dairy cows. The corpus luteum that was diagnosed with false positive or false negative was the developing or regressing states. Thus, ultrasonography was required a serial examination of two or three times accurately diagnosing these corpus luteum.

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