• Title/Summary/Keyword: Porcine

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Characterization of G9 genotype porcine rotavirus isolated in Korea (국내 양돈장에서 분리한 G9형 돼지로타바이러스의 특성)

  • Wang, Jun-Hui;Lee, Seung-Chul;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.35 no.2
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    • pp.83-90
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    • 2012
  • Porcine rotaviruses are the most common causes of viral gastroenteritis in piglets around the world. The major G genotypes of porcine rotaviruses causing diarrhea were G4, G5 and G11 genotypes. Recently, G9 genotype rotaviruses were problemed at swine farms and frequently recognized from diarrheic piglets. In this study, a porcine rotavirus (PoRV-1) was isolated from piglet showing diarrhea using MA104 cells and confirmed as rotavirus by electron microscopy, genomic RNA electropherotyping and indirect immunofluorescence antibody tests. The nucleotide sequence of the VP7 gene of PoRV-1 was determined and compared with those of other genotype rotavirus strains from other parts of the world. Also, the nucleotide sequences of VP4, VP6 and NSP4 genes of PoRV-1 were determined and compared with those of other rotavirus strains from other countries. The results showed that the PoRV-1 isolate belonged to the G9 genotype and the P, I and E genotypes of PoRV-1 were P[23], I5 and E1, respectively. The Korean G9 PoRV-1 isolate and its nucleotide sequence data would be usefully used for the development of porcine rotavirus vaccines in near future.

Localization of Autophagosome in Porcine Follicular Cumulus-oocyte Complex

  • Lee, Seunghoon;Kim, Dong-Hoon;Im, Gi-Sun;Ock, Sun-A;Ullah, Imran;Hur, Tai-Young
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.105-109
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    • 2017
  • Autophagy is an intracellular degradation and recycling system. Oocyte maturation is dynamic process, in which various proteins should be synthesized and degraded. In our previous study, we reported the loci of autophagosome and dynamics of autophagic activity in porcine oocytes during in vitro maturation. In this study, we verified loci of autophagosome in porcine follicular cumulus-oocyte complex by detection of microtubule-associated protein 1A/1B-light chain 3 (LC3) which is the reliable marker of autophagosome. Porcine ovary including various sizes of follicles was fixed within 1 hour after collection from slaughterhouse. After fixation, immunohistochemistry was conducted on sliced ovary tissue containing various sizes of follicles by using LC3 antibody. As a result, LC3 signal was clearly detected in both cumulus and oocytes of various sizes of follicles. We also found ring shaped signal which represent autophagosome near oocyte membrane. Most of the signals in oocytes were localized nearby cellular membrane while evenly dispersed in cumulus cells. Therefore, this result suggests that autophagy occurs in porcine COCs (cumulus-oocyte complexes) at follicular stage.

The Effect of Storage Conditions on the Permeability of Porcine Buccal Mucosa

  • Lee, Jae-Hwi;Lww, Sang-Kll;Choi, Young-Wook
    • Archives of Pharmacal Research
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    • v.25 no.4
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    • pp.546-549
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    • 2002
  • The impact of storage conditions on the permeability of porcine buccal mucosa to [$^3H$]water and [$^{14}C$]mannitol was assessed. The fresh pocine buccal tissue (fresh tissue) was obtained by utilizing pig heads within 24 hours of slaughter. The stored and frozen porcine buccal tissues (stored tissue and frozen tissue) were obtained after the storage of the tissue intact in the pig heads at $4^{\circ}C$ or -$20^{\circ}C$, respectively, for 24 h. The results demonstrated that the barrier properties of the porcine buccal mucosa were maintained with regard to [$^3H$]water permeability when stored at $4^{\circ}C$ for 24 h. However, freezing the tissue resulted in tissue damage illustrated by a significant increase in [$^3$H]water permeability. [$^{14}C$]Mannitol does not appear to be a suitable model solute to assess the ex vivo permeability of porcine buccal mucosa due to its extremely low permeability.

DNA damage repair is suppressed in porcine aged oocytes

  • Lin, Tao;Sun, Ling;Lee, Jae Eun;Kim, So Yeon;Jin, Dong Il
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.984-997
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    • 2021
  • This study sought to evaluate DNA damage and repair in porcine postovulatory aged oocytes. The DNA damage response, which was assessed by H2A.X expression, increased in porcine aged oocytes over time. However, the aged oocytes exhibited a significant decrease in the expression of RAD51, which reflects the DNA damage repair capacity. Further experiments suggested that the DNA repair ability was suppressed by the downregulation of genes involved in the homologous recombination (HR) and nonhomologous end-joining (NHEJ) pathways. The expression levels of the cell cycle checkpoint genes, CHEK1 and CHEK2, were upregulated in porcine aged oocytes in response to induced DNA damage. Immunofluorescence results revealed that the expression level of H3K79me2 was significantly lower in porcine aged oocytes than in control oocytes. In addition, embryo quality was significantly reduced in aged oocytes, as assessed by measuring the cell proliferation capacity. Our results provide evidence that DNA damage is increased and the DNA repair ability is suppressed in porcine aged oocytes. These findings increase our understanding of the events that occur during postovulatory oocyte aging.

Antioxidant Effect of Edaravone on the Development of Preimplantation Porcine Embryos against Hydrogen Peroxide-Induced Oxidative Stress

  • Do, Geon-Yeop;Kim, Jin-Woo;Chae, Sung-Kyu;Ahn, Jae-Hyun;Park, Hyo-Jin;Park, Jae-Young;Yang, Seul-Gi;Koo, Deog-Bon
    • Journal of Embryo Transfer
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    • v.30 no.4
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    • pp.289-298
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    • 2015
  • Edaravone (Eda) is a potent scavenger of inhibiting free radicals including hydroxyl radicals ($H_2O_2$). Reactive oxygen species (ROS) such as $H_2O_2$ can alter most kinds of cellular molecules such as lipids, proteins and nucleic acids, cellular apoptosis. In addition, oxidative stress from over-production of ROS is involved in the defective embryo development of porcine. Previous study reported that Eda has protective effects against oxidative stress-like cellular damage. However, the effect of Eda on the preimplantation porcine embryos development under oxidative stress is unclear. Therefore, in this study, the effects of Eda on blastocyst development, expression levels of ROS, and apoptotic index were first investigated in preimplantation porcine embryos. After in vitro fertilization, porcine embryos were cultured for 6 days in PZM medium with Eda ($10{\mu}M$), $H_2O_2$ ($200{\mu}M$), and Eda+$H_2O_2$ treated group, respectively. Rate of blastocyst development was significantly increased (P<0.05) in the Eda treated group compared with only $H_2O_2$ treated group. And, we measured intracellular levels of ROS by DCF-DA staining methods and investigated numbers of apoptotic nuclei by TUNEL assay analysis is in porcine blastocyst, respectively. Both intracellular ROS levels and the numbers of apoptotic nucleic were significantly decreased (P<0.05) in porcine blastocysts cultured with Eda ($10{\mu}M$). More over, the total cell number of blastocysts were significantly increased (P<0.05) in the Eda-treated group compared with untreated group and the only $H_2O_2$ treated group. Based on the results, Eda was related to regulate as antioxidant-like function according to the reducing ROS levels during preimplantation periods. Also, Eda is beneficial for developmental competence and preimplantation quality of porcine embryos. Therefore, we concluded that Eda has protective effect to ROS derived apoptotic stress in preimplantation porcine embryos.

Activation of Porcine Oocytes Following Intracytoplasmic Injection of Various Sperm Components and foreign species spermatozoa (여러 가지 정자구성성분 및 이종정자 주입에 의한 돼지난자의 활성)

  • Jun, S.H.;Shin, J.S.;Do, J.T.;Kwon, J.K.;Kim, N.H.;Lee, H.T.;Chung, K.S.
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.3
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    • pp.331-340
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    • 1998
  • We determined the incidence of activation, male pronuclear formation and apposition of pronuclei in porcine oocytes following intracy-toplasmic injection of various porcine sperm components and foreign species spermatozoa, such as mouse, human or cattle. The porcine oocytes were activated by injection of a spermatozoon or an isolated sperm head. Neither isolated sperm tail nor perinuclear material removed sperm head activated oocytes. Because injection of mouse, bovine or human spermatozoon activated porcine oocytes, the sperm born activation factors is not strict species specific. Male pronuclear formation and pronuclear apposition were observed in the porcine oocytes following injection of porcine, bovine, mouse or human spermatozoa. The electrical stimulation following sperm cell injection did not enhance the incidence of male pronuclear formation nor pronuclear apposition comparent with sperm cell injection alone (p>0.1). Mitosis and two cell division in some oocytes were observed at 20 to 24 h after injection of porcine spermatozoon. However, none of oocytes following injection of mouse, bovine or human spermatozoa developed to the mitotic metaphase or normally divided to the two cell stage. These results suggested that the oocyte activating factor(s) presented in the perinuclear material and it is not species specific for the porcine oocyte.

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Capacitation and acrosome reaction differences of bovine, mouse and porcine spermatozoa in responsiveness to estrogenic compounds

  • Ryu, Do-Yeal;Kim, Ye-Ji;Lee, June-Sub;Rahman, Md. Saidur;Kwon, Woo-Sung;Yoon, Sung-Jae;Pang, Myung-Geol
    • Journal of Animal Science and Technology
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    • v.56 no.7
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    • pp.26.1-26.10
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    • 2014
  • Background: Endocrine disruptors are exogenous substance, interfere with the endocrine system, and disrupt hormonal functions. However, the effect of endocrine disruptors in different species has not yet been elucidated. Therefore, we investigated the possible effects of $17{\beta}$-estradiol (E2), progesterone (P4), genistein (GEN) and 4-tert-octylphenol (OP), on capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. In this in vitro trial, spermatozoa were incubated with $0.001-100{\mu}M$ of each chemical either 15 or 30 min and then assessed capacitation status using chlortetracycline staining. Results: E2 significantly increased capacitation and the acrosome reaction after 30 min, while the acrosome reaction after 15 min incubation in mouse spermatozoa. Simultaneously, capacitation and the acrosome reaction were induced after 15 and 30 min incubation in porcine spermatozoa, respectively. Capacitation was increased in porcine spermatozoa after 15 min incubation at the lowest concentration, while the acrosome reaction was increased in mouse spermatozoa after 30 min (P < 0.05). E2 significantly increased the acrosome reaction in porcine spermatozoa, but only at the highest concentration examined (P < 0.05). P4 significantly increased the acrosome reaction in bovine and mouse spermatozoa treated for 15 min (P < 0.05). The same treatment significantly increased capacitation in porcine spermatozoa (P < 0.05). P4 significantly increased capacitation in mouse spermatozoa treated for 30 min (P < 0.05). GEN significantly increased the acrosome reaction in porcine spermatozoa treated for 15 and 30 min and in mouse spermatozoa treated for 30 min (P < 0.05). OP significantly increased the acrosome reaction in mouse spermatozoa after 15 min (P < 0.05). Besides, when spermatozoa were incubated for 30 min, capacitation and the acrosome reaction were higher than 15 min incubation in E2 or GEN. Furthermore, the responsiveness of bovine, mouse and porcine spermatozoa to each chemical differed. Conclusions: In conclusion, all chemicals studied effectively increased capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. Also we found that both E2 and P4 were more potent than environmental estrogens in altering sperm function. Porcine and mouse spermatozoa were more responsive than bovine spermatozoa.

Comparison of Microbiological Safety of Porcine Grafts on Gamma Irradiation for Use of Xenografts (돼지 유래 생체 조직의 이식재 활용을 위한 방사선 조사 미생물 제어 평가)

  • Jo, Eu-Ri;Kim, Jeongsoo;Choi, Jong-il;Kim, Jae-Hun;Sung, Nak-Yun;Song, Beom-Seok;Kim, JaeKyung;Park, Jong-Heum;Lee, Ju-Woon
    • Journal of Radiation Industry
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    • v.5 no.3
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    • pp.279-283
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    • 2011
  • This study was compared microbiological safety with gamma-irradiated porcine tendon and skin, as materials for the development of xenografts to regenerate damaged tissues and protect secondary contamination. The porcine tendon and skin were gamma-irradiated after inoculation of bacteria and virus to evaluate irradiation sensitivity of microorganisms. The result showed that the porcine tendon and skin were not different on the sensitivity of microorganisms by gamma irradiation. Bacteria inoculated in the porcine tendon and skin were confirmed that E. coli was the $D_{10}$ values of $0.32{\pm}0.082$ and $0.25{\pm}0.1kGy$ on tendon and skin, and B. subtilis was $4.00{\pm}0.312$ and $3.88{\pm}0.3kGy$ on gamma irradiation, respectively. Moreover, Virus inoculated in the porcine tendon and skin was observed that poliovirus (PV) was $6.26{\pm}0.332$ and $6.88{\pm}0.3kGy$, and porcine parvovirus (PPV) was $1.75{\pm}0.131$ and $1.73{\pm}0.2kGy$ and bovine viral diarrhoea virus (BVDV) was $3.70{\pm}0.212$ and $3.81{\pm}0.2kGy$ on gamma irradiation, respectively. Virus showed higher resistance compared to bacteria on gamma irradiation, but was not detected CPE (cytopathic effect) by virus both tendon and skin at 25 kGy, a standard dose recommended from IAEA for sterilization of medical products. Therefore, These results were considered that gamma irradiation could control effectively bacteria and virus to develop safe porcine xenograft, and apply same irradiation doses to all tissues including tendon and skin of porcine.

Apoptosis Event of Pre-implantation Development Stages in Porcine IVF Embryos (착상 전 돼지 체외수정 배아 발달 단계에서의 세포 자멸사 현상)

  • Hong, Seong-Min;Jeon, Yu-Byeol;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.24 no.3
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    • pp.183-187
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    • 2009
  • In this study, we aimed to determine whether the evaluated markers of cell death could be found at particular developmental stages of normal porcine in vitro fertilization (IVF) embryos. We investigated the characteristics of spontaneous and induced apoptosis during preimplantation development stages of porcine IVF embryos. In experiment 1, to induce apoptosis of porcine IVF embryos, porcine IVF embryos at 22h post insemination were treated at different concentration of actinomycin D (0, 5, 50 and 500 ng/ml in NCSU medium). Treated embryos were incubated at $39^{\circ}C$ in 5% $CO_2$, 5% $O_2$ for 8h, and then washed to NCSU medium and incubated until blastocyst (BL) stage. We examined cleavage rate at 2days and BL development rate at 7days after in vitro culture. A significantly lower rate of cleavage was found in the 500 ng/ml group compared to others (500 ng/ml vs. 0, 5, 50 ng/ml; 27.8 % vs. 50.0%, 41.2%, 35.9%), and BL formation rate in 500 ng/ml was lower than that of others (500 ng/ml vs. 0, 5, 50 ng/ml; 8.0% vs. 12.6%, 11.2%, 12.6%). In experiment 2, to evaluate apoptotic cells, we conducted TUNEL assay based on morphological assessment of nuclei and on detection of specific DNA degradation under fluorescence microscope. This result showed that apoptosis is a normal event during preimplantation development in control group (0 ng/ml actinomycin D). A high number of BL derived control group contained at least one apoptotic cell. Actinomycin D treated BLs responded to the presence of apoptotic inductor by significant decrease in the average number of blastomeres and increase in the incidence of apoptotic cell death. In 500 ng/ml group, the incidence of apoptosis increased at 4-cell stage and later. This result suggested that apoptosis is a process of normal embryonic development and actinomycin D is useful tool for the apoptosis study of porcine preimplantation embryos.

Microbial Transglutaminase Modifies Gel Properties of Porcine Collagen

  • Erwanto, Y.;Kawahara, S.;Katayama, K.;Takenoyama, S.;Fujino, H.;Yamauchi, K.;Morishita, T.;Kai, Y.;Watanabe, S.;Muguruma, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.2
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    • pp.269-276
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    • 2003
  • We studied the gel properties of porcine collagen with microbial transglutaminase (MTGase) as a catalyst. A creep meter was used to measure the mechanical properties of gel. The results showed samples with high concentration of MTGase gelled faster than those with a low concentration of MTGase. The gel strength increased with incubation time and the peaks of breaking strength for 0.1, 0.2 and 0.5% MTGase were obtained at 40, 20 and 10 min incubation time, respectively. According to SDS-PAGE, the MTGase was successfully created a collagen polymer with an increase in molecular weight, whereas no change in formation was shown without MTGase. The sample with 0.5% MTGase began to polymerize after 10 or 20 min incubation at $50^{\circ}C$, and complete polymerization occurred after 40-60 min incubation. Scanning electron microscopic analysis revealed that the gel of porcine collagen in the presence of MTGase produced an extremely well cross-linked network. The differential scanning calorimetric analysis showed the peak thermal transition of porcine collagen gel was at $36^{\circ}C$, and that with MTGase no peak was detected during heating from 20 to $120^{\circ}C$. The melting point of porcine collagen gel could be controlled by MTGase concentration, incubation temperature and protein concentration. Knowledge of the structural and physicochemical properties of porcine collagen gel catalyzed with MTGase could facilitate their use in food products.