• Title/Summary/Keyword: Chicken embryo

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HATCHING OF CULTURED EMBRYOS OF THE PEKING DUCK

  • Li, Z.D.;Qi, S.Z.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.2
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    • pp.195-197
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    • 1996
  • Peking duck embryos incubated for 82 hrs after oviposition, stage 18 to 20(Hamberger and Hamilton, 1951) were transferred into the recipient eggshells and further incubated at $38.0^{\circ}C$ RH(relative humidity) 60 to 70%, with rotation at an angle of $30^{\circ}$ and the frequency of 4 times an hour. The survival rate was 100, 98, 82, 82, 70, 46, 21 and 16%, after the incubation period of 4, 8, 12, 16, 20, 24, 28 and 29 days, respectively. The present result clearly shows that the ex ova culture is possible for the Peking duck just as in the cases of the chicken or the quail. This culture technique could be useful for experimental manipulation of the embryos.

Studies on Properties of Avian Reovirus Isolated in Korea (국내에서 분리한 닭 레오바이러스의 성상에 관한 연구)

  • 김성식;박병옥;김순재
    • Korean Journal of Veterinary Service
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    • v.15 no.1
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    • pp.67-80
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    • 1992
  • Avian reoviruses have been implicated in respiratory disease enteric conditions including Cloacal pasting in young thicks, pericarditis, hydropericardium, anaemia with swollen spleen and liver and petechiation of skeletal muscle and viral arthritis. This study was conducted to examine properties of reovirus field 3 strains isolated from affected broiler from several farms. An infectious agent was isolated from leg tendons and intestine of broiler with clinical tenosynovitis. The agent grew well on the chicken embryo kideny cells(CEK). One of them produced cytopathic effects(CPE) of round type and formed intranuclear inclusions, and the other was characterized by CPE of syncytical type and cytoplasmic inclusion. The properties and serological classification of field strains were examined by hemagglutin test, virus neutralization test, agar gel precipitin, electropherotype. They showed no hemagglutination reactions and not well neutralization and to possess common antigens detectable by AGP test. RNA electropherotype presented 10 segment band as the previous report. These data suggest that the field strains and standard strains (1133, 1733) may be the same group.

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Identification of Gene Locus by the Somatic Cell Hybridization in Chicken (체세포 융합에 의한 닭의 유전인자구명에 관한 연구)

  • 정익정
    • Korean Journal of Poultry Science
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    • v.16 no.1
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    • pp.1-8
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    • 1989
  • This experiment was conducted to improve the performance of chickens by the precise separation and analysis of chromosomes which are integrated genetic materials, and by the use of gene manipulation techniques. Following are the main results obtained. 1. When the chromosomes were separated through the leucocyte culture and analyzed by Giemsa banding techniques (especially by the method in which 20 layers of banding patterns could be found in chromosome #1), the normal Patterns of chromosomes #l-9 and sex chromosomes, and the location of constitutive heterochromatin without any gene activities in all chromosomes were discovered. 2. To utilize the primodial germ cells (PGC) as the genetic vector which is one of the most important gene manipulation techniques, PGC's from triploid were transplanted to normal host embryos. Since the donor PGC's(3n) were found in the gonads of growing host embryos gene manipulation in poultry using PGC's, seemed to be possible.

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Egg Antibody Farming and IgY Technology for Food and Biomedical Applications

  • Sim, J.S.;Sunwoo, H.H.
    • Korean Journal of Poultry Science
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    • v.31 no.1
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    • pp.37-44
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    • 2004
  • It has been recognized that the hen, like its mammalian counterparts, provides young chicks with antibodies as protection against hostile invaders. This system facilitates the transfer of specific antibodies from serum to egg yolk, and provides a supply of antibodies called immunoglobulin Y(IgY) to the developing embryo and the hatched chick. The protection against pathogens that the relatively immune-incompetent newly hatched chick has, is through transmission of antibodies from the mother via the egg. Egg yolk, therefore, can be loaded with a large amount of IgY against pathogens which can immobilize the existing or invading pathogens during the embryo development or in day-old chicks. Thus, the immunization of laying hens to various pathogens results in production of different antigen-specific IgY in eggs. Egg yolk contains 8∼20 mg of jmmunoglobulins (IgY) per ml or 136∼340 mg per yolk suggesting that more than 30 g of IgY can be obtained from one immunized hen in a year. By immunizing laying hens with antigens and collecting IgY from egg yolk, low cost antibodies at less than $10 per g compared to more than $20,000 per g of mammalian IgG can be obtained. This IgY technology opens new potential market applications in medicine, public health, veterinary medicine and food safety. A broader use of IgY technology could be applied as biological or diagnostic tool, nutraceutical or functional food development, oral-supplementation for prophylaxis, and as pathogen-specific antimicrobial agents for infectious disease control. This paper has emphasized that when IgY-loaded chicken eggs are produced and consumed, the specific antibody binds, immobilizes and consequently reduces or inhibits the growth or colony forming abilities of microbial pathogens. This concept could serve as an alternative agent to replace the use of antibiotics, since today, more and more antibiotics are less effective in the treatment of infections, due to the emergence of drug-resistant bacteria.

Molecular Sexing and Species Identification of the Processed Meat and Sausages of Horse, Cattle and Pig

  • Kim, Yoo-Kyung;Kang, Yong-Jun;Kang, Geun-Ho;Seong, Pil-Nam;Kim, Jin-Hyoung;Park, Beom-Young;Cho, Sang-Rae;Jeong, Dong Kee;Oh, Hong-Shik;Cho, In-Cheol;Han, Sang-Hyun
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.61-64
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    • 2016
  • We developed a polymerase chain reaction (PCR)-based molecular method for sexing and identification using sexual dimorphism between the Zinc Finger-X and -Y (ZFX-ZFY) gene and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for mitochondrial DNA (mtDNA) cytochrome B (CYTB) gene in meat pieces and commercial sausages from animals of different origins. Sexual dimorphism based on the presence or absence of SINE-like sequence between ZFX and ZFY genes showed distinguishable band patterns between male and female DNA samples and were easily detected by PCR analyses. Male DNA had two PCR products appearing as distinct two bands (ZFX and ZFY), and female DNA had a single band (ZFX). Molecular identification was carried out using PCR-RFLP of CYTB gene, and showed clear species classification results. The results yielded identical information on the sexes and the species of the meat samples collected from providers without any records. The analyses for DNA isolated from commercial sausage showed that pig was the major source but several sausages originated from chicken and Atlantic cod. Applying this PCR-based molecular method was useful and yielded clear sex information and identified the species of various tissue samples originating from livestock.

Recolonization of Transfected Blastodermal Cells in Developing Embryos after Transferring into UV-irradiated Fertilized Hen′s Egg (UV-조사 수정란 내로 이식한 유전자 변화 배반엽 세포의 재구성)

  • Lee, K.S.;Lee, H.;Kim, K.D.;Park, S.S.;Lee, S.H.
    • Korean Journal of Poultry Science
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    • v.27 no.2
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    • pp.155-161
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    • 2000
  • Unfortunately, there is no technique which is stable and repetitive to produce transgenic chicken, although various ways of gene transfer including PGC-and embryonic cell-mediated gene transfer, DNA microinjection, virus inoculation and sperm cells have been employed. The aims of this study were 세 develop and establish such a stable, repetitive and efficient way of gene transfer giving a faithful gene expression during development after the reconstruction of embryo in an UV-irradiated egg. A dual reporter plasmid (pJJ9), a fusion gene containing lacZ and GFP driven by a CMV promoter was used to exploit either merits of both reporting markers. lacZ with strong signal or GFP with vital marking. Electroporated embryonic blastodermal cells (EBCs) in the presence of the pJJ9 DNA faithfully showed 377 bp PCR product and lacZ or GFP expressions in the identical cells in vitro of in vivo. Furthermore, analyses of expression pattern of the foreign DNA demonstrated that microinjected EBCs cells into the UV-irradiated recipient egg should participate in normal developmental process, for example, proliferation and differentiation into various tissues. Thirty percentages of the manipulated eggs showed lacZ expression in their tissues. These results together with the specific procedures used in this study should facilitate avian transgenesis.

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The Expression Patterns of Cdc25A, Cdc25B, Sox2 and Mnb in Central Nervous System in Early Chicken Embryos

  • Zhang, Hui;Qin, Junhui;Cao, Jingjing;Hei, Nainan;Xu, Chunsheng;Yang, Ping;Liu, Haili;Chu, Xiaohong;Bao, Huijun;Chen, Qiusheng
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.6
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    • pp.781-787
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    • 2009
  • The sense and antisense digoxigenin-labeled RNA probes of four genes, Cdc25A, Cdc25B, Sox2 and Mnb, were produced by using SP6 and T7 RNA polymerases, respectively, and in vitro transcription. Expression patterns of the four genes were detected by in situ hybridization in HH (Hamburger and Hamilton) stage 10 chick embryos. In general, expression patterns of the four genes were similar. mRNA of the four genes was mostly restricted to the entire CNS (central nervous system). All were confined to an identical region, neural tube, neural groove and caudal neural plate, corresponding to the notochord or spinal cord, but there was some distinction in specific region or in concentration, for example in somites. The overlap in expression at the same developmental stage in the CNS suggests that the four genes may be functional similar or related in CNS development. Expression patterns of the four genes support specific roles of these regulators in the developing CNS.

EFFECT OF LIPOPOLYSACCHARIDE AND INTERFERON-${\gamma}$ ON THE FORMATION OF OSTEOCLAST-LIKE MULTINUCLEATED CELL FROM CHICKEN BONE MARROW CELLS IN VITRO (세포 배양시 닭 골수세포로부터 파골세포앙 세포형성에 지질다당류와 인터페론 감마가 미치는 영향)

  • Oh, Hong-Kyun;Kim, Jung-Keun;Lee, Jae-Hyun
    • Journal of Periodontal and Implant Science
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    • v.25 no.3
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    • pp.659-667
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    • 1995
  • 파골세포는 조혈기관 단핵의 세포로부터 생성되어 골 홉수에 중요한 역할올 담당하며, 지질다당류는 그람음성균의 세포벽을 이루는 성분으로서 치주질환시 치조골 홉수에 관여한다고 알려져 왔다. 활성화된 림프구, 대식세포와 단핵세포로부터 생성되는 당단백질인 인터페론 감마는 파골세포에 의한 골홉수를 억제한다고 밝혀졌다. 이 연구 논문의 목적은 지질다당류와 인터페론 감마가 닭 골수의 미분화세포가 파골양세포로 전환되는데 어떠한 영향올 주는지를 알아보기 위함이다. 16${\sim}$18 일째의 닭의 배 (chick embryo) 에서 경골을 분리하고 횡절개하여 혈청없는 M-199 배양액에 보관했다. 이것을 9${\mu}m$ filter로 여과시켜서 이미 분화된 파골세포와 기타 다른 분화 세포를 분리했다. 여기에서 파골세포의 전구세포를 얻어 LPS와 IFN-${\gamma}$를 단독 또는 복합처리 하고나서 4일 후에 tartrate resistant acid phosphatase (TRAP) Stain을 시행하고 TRAP 양성이며 핵이 세개 이상인 다핵의 세포형성을 관찰하여 세포를 계수하여 다음과 같은 결과를 얻었다. 1. 닭에서 분리해낸 미분화세포에 0.1. 0.5. 1.0 ${\mu}/ml$ 의 LPS 농도를 처리하고 1 주일간 배양한 결과. 0.1 ${\mu}/ml$ 의 농도에서는 대조군에 비해 TRAP 양성인 파골양세포가 증가하는 경향을 보였으며, 반면에 LPS는 0.5 와 1.0 ${\mu}/ml$ 의 농도에서 세포독성을 보였다.(P<0.05) 2. IFN-${\gamma}$는 50. 500U/ml 의 농도에서 대조군에 비해 TRAP 양성인 파골양세포의 수가 감소하는 경향올 보였다 .3. INF-${\gamma}$는 LPS 에의해 유도된 TRAP 양성인 파골양세포의 형성을 감소시켰고 특히 . 250.500U/ml 의 농도에서 유의 성 있는 감소를 보였다. 위의 결과로부터 LPS는 닭의 골수세포로부터 파골양세포의 형성을 증가시키며 IFN-${\gamma}$는 LPS에의해 유도된 파골양세포수를 감소시킨다는 결론을 얻었다.

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Standardization of an enzyme-linked immunosorbent assay for detection of antibody to avian reticuloendotheliosis virus (세망내피증 바이러스 항체검출을 위한 ELISA 표준화)

  • Sung, Haan Woo;Lee, Su Jeong
    • Korean Journal of Veterinary Research
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    • v.45 no.4
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    • pp.569-574
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    • 2005
  • Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.

Exper mental Studies of Fowl Pox Vaccine (계두예방약(鷄痘豫防藥)의 실험적연구(實驗的硏究))

  • Im, Yeong-Mun;Byeon, Eun-Su
    • Korean Journal of Veterinary Research
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    • v.1 no.1
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    • pp.36-53
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    • 1961
  • This experiment was conducted on the fowl pox embryo vaccine for the production immunity, and stability, using an attenuated fowl pox virus (Nakano strin). Burnet's window method was applied, that is, 0.1 ml. of seed virus was inoculated on the chorioallantoic membrane of 12-day old chicken embryos, and incubated for 5 to 6 day, and then the result were read. Four kinds of suspensions of different embyo tissue were prepared and tested for the infectivity in chickens. Finally the suspension of chorioallantoic membrane was used as the vaccine throughout the experiment. Results obtained in this experiment are summarized as follows: (1) Of embryo tissue infected with the vaccine virus, chorioallantoic membrane had the highest virus titer of $10^{-5.4}$ $EID_{50}$, and albumen the lowest titer of $10^{-0.7}$ $EID_{50}$. (2) Suspensions of infected whole embryo with or without saline, and de-embryonated whole egg had about the same virus titer of $10^{-4.4}$ $EID_{50}$, whereas the chorioallantoic membrane had $10^{-5.7}$ EID 50 or higher. The virus titer droped one log from $EID_{50}$ when inoculated into chickens. Takes were observed 35.6% of 500 chickens by stick method and 89% of 500 chickens by brush method. (3) The chorioallantoic membrane conferred almost perfect immunity for chickens by 10 days after vaccination. (4) Satisfactory immunity was observed in the chickens when eruption in a single follicle. (5) Eight of 10 vaccinated chickens revealed durable immunity for 307 days following vaccination. (6) The vacuum-dried vaccine maintained its infectiviy for 899 days at $5^{\circ}C$ or below and maintained the vius titer of $10^{-3.6}$ $EID_{50}$. On the other hand, non-desiccated wet vaccine maintained the titer of $10^{-3.0}$ $EID_{50}$ for 50 days of preservation period at $5^{\circ}$. However, in 50% glycerin-saline the infectivtiy of the same wet vaccine dropped to $10^{-1.5}$ $EID_{50}$ (7) The vartation of virus titer of the vaccine before and after desiccation was $10^{-0.5}$ $EID_{50}$ on the average. (8) As suspending media, 0.85 per cent saline and distilled water showed nearly the same effect on the infectivity of the vaccine by retaining the titer $10^{-3.0}$ $EID_{50}$ after 50 days of preservation both at $5^{\circ}C$ and $20^{\circ}C$, while 50 percent cent glycerine-saline dropped the titer to $10^{-2.5}$ EID and $10^{-1.5}$ $EID_{50}$ respectively at $5^{\circ}C$ and $2^{\circ}C$ after the same period.

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