• Title/Summary/Keyword: Porcine

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Development of oligonucleotide microarray system for differential diagnosis of enteric viruses in diarrheic fecal samples in pigs

  • Park, Nam-Yong;Kim, Yong-Hwan;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.30 no.4
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    • pp.489-496
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    • 2007
  • An oligonucleotide microarray system was developed for the simultaneous detection of porcine epidemic diarrhea virus, transmissible gastroenteritis virus, porcine enteric calicivirus, porcine group A and C rotavirus. RNAs of the reference viruses and porcine diarrhea samples were extracted and amplified using one-step multiplex RT-PCR in the presence of cyanine 5-dCTP and hybridized on the microarray chip that spotted the virus-specific oligonucleotides. This system were approximately 10-to 100-fold higher in sensitivity than conventional RT-PCR, and the assay time was less than 3 hours. The relative sensitivity and specificity were 92% and 72.2%, respectively, based on 102 porcine diarrhea samples using RT-PCR as gold standard. These results suggested that the oligonucleotide microarray system in this study be probably more reliable and reproducible means for detecting porcine enteric viruses and that it could be of substantial use in routine diagnostic laboratories.

Cloning, Sequencing and Expression Analysis of Porcine Uroplakin II Gene

  • Gwon Deuk-Nam;Kim Jin-Hoe
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.90-90
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    • 2002
  • In this study, we report the cloning of the porcine UPII genomic DNA, which contains a putative full-length open reading frame encoding the UPII protein. A comparison of the porcine UPII gene coding sequence with the previously published mouse UPII sequence demonstrates that only the exon sequences are partially conserved. Northern and immunohistochemical analyses show that the porcine UPII gene is expressed only in the urothelium and that the protein specifically localizes to urothelial superficial cells. (omitted)

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Effect of Co-culture with Spermatozoa on the Resumption of Meiosis in Porcine Germinal Vesicle Oocytes Arrested with Meiotic Inhibitors

  • Kim, Byung-Ki
    • Reproductive and Developmental Biology
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    • v.29 no.4
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    • pp.223-227
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    • 2005
  • In vitro maturation of porcine immature cumulus-enclosed oocytes can be enhanced by co-incubation with spermatozoa even before fertilization. The aim of this study was to determine whether the addition of spermatozoa into the culture medium can stimulate the meiosis resumption of porcine cumulus-enclosed oocytes arrested at germinal vesicle (GV). Cumulus-enclosed oocytes (CEOs) were collected from follicles of 3 to 5mm diameter. Porcine CEOs were cultured in tissue culture medium containing various meiosis inhibitors and spermatozoa. Oocytes were examined for evidence of GV and GV breakdown after 24 h culture. After 24 h culture $43.8\%$ of oocytes cultured in only TCM 199 remained at GV stage whereas $56.2\%$ of oocytes were able to resume meiosis. When porcine CEOs were cultured in the medium with meiosis inhibitor such as, dibutyryl cAMP (dbcAMP) and forskolin (Fo), more than $90\%$ of oocytes were not able to resume meiosis. However, co-culture of porcine CEOs with spermatozoa was able to overcome the inhibitory effect of dbcAMP and Fo. Irrespective of the presence of 3-isobutyl-1-methylxanthine (IBMX), no difference was observed in the proportion of oocyte reached germinal vesicle breakdown (GVBD). The present study suggests that dbcAMP and Fo prevent the spontaneous maturation of competent oocyte in culture after isolation from follicles and that mammalian spermatozoa contain a substance(s) that improves meiosis resumption in vitro of porcine cumulus-enclosed oocytes.

Development of a Chemically Defined In Vitro Maturation System for Porcine Oocytes: Application for Somatic Cell Nuclear Transfer

  • Koo, Ja-Min;Won, Cheol-Hee;Min, Byung-Moo;Roh, Sang-Ho
    • International Journal of Oral Biology
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    • v.30 no.4
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    • pp.131-134
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    • 2005
  • In the present study, performances of several in vitro maturation (IVM) systems for porcine follicular oocytes were evaluated, and an efficient chemically defined IVM system for porcine oocytes was proposed. The proposed one-step culture system supplemented with polyvinylalcohol (PVA) gave competitive efficiencies in terms of oocyte maturation and blastocyst development after parthenogenetic activation and in vitro culture, compared with the conventional two-step culture system by a supplementation of porcine follicular fluid (pFF). Additionally, it is identified that the proposed chemically defined one-step culture system yielded the comparable level of blastocyst production to the conventional maturation system in porcine somatic cell nuclear transfer (SCNT). Therefore, one can eliminate un-expected effects accompanied by supplementation of pFF. No medium replacement during whole maturation period is an additional benefit by applying this new system. Thus, these data support that the developed PVA supplemented chemically defined one-step IVM system for porcine follicular oocyte might be used in porcine SCNT program.

Detection of viral pathogens and isolation of porcine circovirus 2 from postweaning multisystemic wasting syndrome-affected piglets (이유자돈 전신소모성증후군 이환 자돈에서의 바이러스성 원인체 검색 및 porcine circovirus 2 분리동정)

  • Park, Choi-Kyu;Kim, Hyun-Soo
    • Korean Journal of Veterinary Research
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    • v.44 no.4
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    • pp.561-569
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    • 2004
  • To detect viral agents and isolate porcine circovirus 2 (PCV2), 60 samples of lung and lymph node were collected from 5 to 12 week-old pigs that had showed clinical signs of postweaning multisystemic wasting syndrome (PMWS). Polymerase chain reactions (PCRs) were conducted to identify the viral pathogens including PCV1, PCV2, porcine parvovirus (PPV) and porcine reproductive and respiratory syndrome virus (PRRSV) that have been considered to be the causal agents of PMWS. Among 60 samples, PCV 2 was detected from 57 samples but no PCV 1 was detected. PRRSV and/or PPV were also detected from 27 (47.4%) samples and 1 (1.8%) sample of these 57 PCV 2-positive samples, respectively. Tissue homogenates were inoculated onto PCV-free PK-15 cell monolayers. Seven isolates were confirmed as PCV 2 by multiplex PCR, indirect immunofluorescence assay, and transmissible electron microscopy. These date suggest that PRRSV is a major cofactors causing PMWS in pigs that were infected with PCV2 in Korea.

Comparative scanning electron microscope analysis of the enamel of permanent human, bovine and porcine teeth

  • Olek, Aneta;Klimek, Leszek;Boltacz-Rzepkowska, Elzbieta
    • Journal of Veterinary Science
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    • v.21 no.6
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    • pp.83.1-83.14
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    • 2020
  • Background: Bovine and porcine teeth are often used in in vitro experiments as substitutes of human teeth. Objectives: The aim of the present study was to perform a comparative analysis of enamel morphology of permanent human, bovine and porcine teeth under the scanning electron microscope. Methods: As many as 10 human, 10 bovine, and 10 porcine teeth were studied. All the teeth were sectioned and the halves were randomly divided into 2 groups according to the examined tissue (vestibular enamel at the mid-height of the dental crown and in the cervical area). Human and bovine enamel was etched for 15 sec and porcine enamel for 30 sec. The scanning electron microscope analysis was performed. The length and width of enamel prisms were determined with the "Met-Ilo" 1.1 computer program. Results: All enamel samples revealed the same etching pattern-Silverstone's type 2. Bovine enamel showed a similar porosity and the amount of interprismatic enamel compared to human enamel while the amount and width of interprismatic enamel bands in porcine enamel were evidently greater. The shape of the porcine prisms was visually similar to human prisms, although dimensions were significantly different. However, bovine prisms differed in form and appeared to be distinctly elongated. Conclusions: Reported findings indicate that the results of experimental studies carried out on bovine and porcine enamel should not be compared with the results obtained on human enamel.

Expression of the Antioxidant Enzyme and Apoptosis Genes in in vitro Maturation lin vitro Fertilization of Porcine Embryos

  • H. Y. Jang;H. S. Kong;Park, K. D.;G. J. Jeon;Lee, H. K.;B. K. Yang
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.47-47
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    • 2003
  • The present study was conducted to determine the expression of the antioxidant enzyme(CuZn-SOD, Mn-SOD and GPX and apoptosis gene(caspase-3) for in vitro culture in in vitro maturation and in vitro fertilization(IVM/IVF) embryos in porcine. Porcine embryos derived from IVM/IVF were cultured in NCSU23 medium under 5% $CO_2$ in air at 38.5$^{\circ}C$. The patterns of gene expression for several antioxidant enzyme and apoptosis genes during preimplantion porcine embryo development were examined by the modified semi-quantitative single cell reverse transcriptase- polymerase chain reaction (RT-PCR). Preimplantation porcine embryos produced by IVM/IVF have expressed mRNAs for CuZn-SOD and GPX, whereas transcripts for Mn-SOD have not detected at any developmental stages. Expression of caspase-3 mRNA was detected at 2 cell, 8 cell, 16 cell and morula stages. The fas ligand transcripts were detected in porcine blastocyst. These results suggest that various antioxidant enzymes and apoptosis genes play crucial roles in in vitro culture of porcine IVM/IVF embryos.

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Effects of in vitro Culture Period of Reconstructed Embryos and Genetic Background of Feeder Cells on Establishment of Embryonic Stem Cells Derived from Somatic Cell Nuclear Transfer Blastocysts in Pigs

  • Han, Na Rae;Baek, Song;Lee, Yongjin;Lee, Joohyeong;Yun, Jung Im;Lee, Eunsong;Lee, Seung Tae
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.1
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    • pp.86-93
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    • 2020
  • The establishment of porcine embryonic stem cells (ESCs) from porcine somatic cell nuclear transfer (SCNT) blastocysts is influenced by in vitro culture day of porcine reconstructed embryo and feeder cell type. Therefore, the objective of the present study was to determine the optimal in vitro culture period for reconstructed porcine SCNT embryos and mouse embryonic fibroblast (MEF) feeder cell type for enhancing colony formation efficiency from the inner cell mass (ICM) of porcine SCNT blastocysts and their outgrowth. As the results, porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days showed significantly increased efficiency in the formation of colonies, compared to those for 7 days. Moreover, MEF feeder cells derived from outbred ICR mice showed numerically the highest efficiency of colony formation in blastocysts produced through in vitro culture of porcine SCNT embryos for 8 days and porcine ESCs with typical ESC morphology were maintained more successfully over Passage 2 on outbred ICR mice-derived MEF feeder cells than on MEF feeder cells derived from inbred C57BL/6 and hybrid B6CBAF1 mice. Overall, the harmonization of porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days and MEF feeder cells derived from outbred ICR mice will greatly contribute to the successful establishment of ESCs derived from porcine SCNT blastocysts.

Effects of Glucose, Lactate and Pyruvate on Development of In Vitro Matured and Fertilized Porcine Embryos (Glucose, Lactate 및 Pyruvate가 돼지 체외수정란의 초기발생능에 미치는 영향)

  • 오건봉;박병권;서길웅;이규승
    • Korean Journal of Animal Reproduction
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    • v.19 no.1
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    • pp.9-14
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    • 1995
  • This study was conducted to investigate the effect of energy source on development of in vitro development of in vitro matured and fertilized porcine 2-cell embryos. The relative preferences of glucose, lactate and pyruvate for in vitro development of porcine 2-cell embryos were determined. The results obtained are as follows. 1. 33.3, 20.8 and 29.2% of porcine embryos reached morula stage in addition to lactate, glucose, and both glucose and lactate in the culture medium as energy source, respectively. 2. 38.5, 15.4 and 26.9% of porcine embryos reached morula stage in addition to pyruvate, glucose, and both glucose and pyruvate in culture medium as energy source, respectively. 3. 42.9, 21.4 and 28.6% of porcine embryos reached morula stage in addition to pyruvate and lactate, glucse alone, and glucose, lactate and pyruvate in culture medium as energy source, respectively.

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Factors Affecting In-Vitro Maturation in Porcine Oocytes (돼지난자의 체외성숙에 영향을 미치는 요인)

  • 박춘근
    • Journal of Embryo Transfer
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    • v.11 no.2
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    • pp.179-191
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    • 1996
  • In-vitro culture has provided new inforrnation on mechanisms of oocytes rnaturation and results obtained in vitro have led to new questions. In porcine, follicular and oocyte size have the crucial importance for the oocytes maturation. The addition of hormones to the culture medium was found to accelerate and facilitate meiotic maturation. The presence of some factors in serum trigger the resumption of meiosis and support the maturation of oocytes in vitro. The maturation rate of porcine oocytes was also increased by supplementation of porcine follicular fluid to the culture medium. The growth factors can stimulate nuclear maturation and enhances cytoplasnic maturation of oocytes by interaction with gonadotropins. The maturation-promoting factor brings about GVBD and the subsequent maturational events in oocytes. However, cAMP can block the spontaneous meiotic maturation of oocytes in culture. The understanding of these influences is a prerequisite to enhancing in vitro maturation of porcine oocytes.

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