• Korean Journal of Veterinary Research
• /
• v.41 no.1
• /
• pp.29-35
• /
• 2001

The protective effects of egg yolk atibodies obtained from chickens immunized with fimbrial antigens from ETEC 987P were evaluated in 14 and 21 d old pigs in which ETEC diarrhea was induced. For the Exp. 1, eight early-weaned pigs($5.00{\pm}0.5kg$ average BW and 14 d average age) and eight weaned pigs($6.00{\pm}0.5kg$ average BW and 21 d average age) were used to examine influence of egg yolk antibodies on growth performance and resistance to ETEC 987P infection. Dietary treatments included 1) administered of commercial egg yolk(14 d of age; CEY14), 2) administered of egg yolk antibodies(14 d of age; EYA14), 3) administered of commercial egg yolk(21 d of age; CEY21), 4) administered of egg yolk antibodies(21 d of age; EYA21). The 14 and 21 d old pigs were challenged with 2 ml of ETEC 987P at a dose of $10^{10}\;CFU\;ml^{-1}$ per weaned pigs. Weaned pigs treated with egg yolk antibodies recovered and pigs treated with egg yolk antibodies tended to increase average daily gain(P<0.05). Also, EYA12 and EYA21 treatments were reduced coli-form bacteria concentration and increased Lactobacilli sp. concentration from feces. For the Exp. 2, sixteen weaned pigs($6.00{\pm}0.5kg$ average daily gain BW and 21 d average age) were used to examine influence of yolk or white from egg containing antibodies on growth performance and resistance to ETEC 987P infection. Dietary treatments included l) administered of commercial egg yolk(CEY), 2) administered of commercial egg white(CEW), 3) administered of egg yolk antibodies(EYA), 4) administered of egg white antibodies(EWA). Pigs treated only with EYA showed signs of recovery. Also, EYA treatment showed the best average daily gain without significant differences (P>0.05). EYA treatment was reduced coli-form bacteria concentration increased and Lactobacilli sp. concentration from feces. In conclusion, egg yolk antibodies have protective effects from pigs in which ETEC diarrhea was induced.

• Research in Plant Disease
• /
• v.12 no.2
• /
• pp.144-147
• /
• 2006

Egg yolk immunoglobulin (IgY) is much widely used in medical fields, but its use in serology of plant viruses is much limited. We produced an IgY against pepper mild mottle virus (PMMoV) and applied it to several serological tests. Polyclonal antibodies were obtained from the egg yolk of chicken immunized with a total of 2mg of purified PMMoV over 2 months. The titers of antibodies were measured with the ring-test over six months after the first injection. The highest.titers of IgY was 1/2,560 at 2 months after the first injection. Approximately 60-80 mg of IgY were obtained from one egg yolk. Using the IgY, 1ng/ml of purified PMMoV was detected with the indirect ELISA. Gelrite gel double diffusion test, ELISA and tissue immuno-binding assay employing IgY gave similar sensitivity and specificity to those of IgG developed in rabbit. Therefore, the IgY which can be obtained in large quantities from a chicken, might be useful for the antibody production and the serology of plant viruses.

• Horticultural Science & Technology
• /
• v.32 no.4
• /
• pp.544-549
• /
• 2014

Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf and bulb samples showing characteristic symptoms of virus infection were collected in 2012, and 80 field samples were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The infection frequencies were 79% for LMoV, 5% for LSV, and 3% for CMV. The LMoV coat protein gene was amplified and cloned into the pET21d(+) expression vector to develop serological diagnostic tools to detect LMoV. The resulting carboxy-terminal His-tagged coat proteins were expressed in Escherichia coli strain BL21 (DE3) by induction with IPTG. The recombinant proteins were purified using Ni-NTA agarose beads and used as an antigen to produce polyclonal antibodies in laying hens. The resulting egg yolk immunoglobulin (IgY) specifically recognized LMoV from infected plant tissues in immunoblotting assays and had comparable sensitivity to that of a mammalian antibody. In addition, method of immunocapture RT-PCR using this IgY was developed for sensitive, efficient, and rapid detection of LMoV. Based on these results, large-scale bulb tests and detection of LMoV in epidemiological studies can be performed routinely using this IgY. This is the first report of production of a polyclonal IgY against a plant virus and its use for diagnosis.

• YAKHAK HOEJI
• /
• v.49 no.1
• /
• pp.6-10
• /
• 2005

Immunoglobulin Y (IgY) obtained from chicken as the immunization host brings several advantages to antibody production, such as improved yield, lower cost, longer stability and higher specificity than mammalian immunoglobulin. In the present study, we attempted to produce IgY against a sialic acid-binding lectin, Maackia fauriei agglutinin (MFA), from egg yolk of white Leghorn hens. For the isolation of IgY from egg yolk, we applied a water dilution method. The weekly yield of IgY was determined by enzyme-linked immunosorbent assay, with a final yield of anti-MFA IgY from total IgY of approximately $1\%$. The yielded IgY were used to prepare IgY-affinity column conjugated with CNBr-activated Sepharose 4B, which resulted in the lectin being successfully purified in a single step from Maackia fauriei. This purified lectin exhibited the same hemagglutination activity as lectin purified using conventional purification methods.

• Microbiology and Biotechnology Letters
• /
• v.36 no.4
• /
• pp.299-306
• /
• 2008

Human pancreatic lipase is a digestive enzyme which is synthesized in pancreas, secreted into small intestine, and there hydrolyze the fat in food. Pancreatic lipase protein composes of catalytic domain and colipase-binding domain. In this research, the gene segments corresponding to total protein, catalytic domain, and co lipase-binding domain were cloned by PCR method, inserted into an expression vector, and then used to transform Escherichia coli BL21 (DE3). The recombinant proteins produced were purified and injected intramuscularly three times into laying hens. The egg yolk antibodies (IgY) were obtained from the egg yolks and tested for their antibody titer. Among three IgY, the IgY against colipase-binding domain showed the highest antibody titer. All three IgY had inhibitory effects on the porcine pancreatic lipase. Among them, the IgY against colipase-binding domain showed the highest inhibition effects. The fat diet with corn oil and IgY was administrated to the experimental rats and their blood compositions were examined with time course. The triglyceride concentration of treated rats was decrease meaningfully when compared with those of control rats. This suggested that the IgY against colipase-binding domain antigen inhibited pancreatic lipase in vivo.

• Food Science of Animal Resources
• /
• v.32 no.6
• /
• pp.706-712
• /
• 2012

The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 ${\mu}g/mL$, having ca. $10^8$ coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of $10^5$ CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

• Korean Journal of Veterinary Research
• /
• v.44 no.3
• /
• pp.415-420
• /
• 2004

Immunoprophylatic effect of IgY against B. bronchispetica was proven with 100% preventive rate in mice administrated with IgY with antibody titer 1:640~1:2,560. Intramuscular administration was more efficient than oral administration. This phenomenon was also observed in the therapeutic effects of IgY after challenge with B. bronchseptica in mice. In the field trials with the egg yolk antibodies from hens immunized with combined antigens with B. bronchiseptica and parvovirus, curing rates in dogs with severe clnical signs such as bloody diarrhea were 81.6% and 86.7% by intramuscular or subcutaneous administration of IgY, respectively. Safety of the antibodies in dogs was proven without any side effects such as vomiting, edema, fever, etc. by adminstration of double doses for 7 days. These results indicated that the egg yolk antibodies could be used as effective prevention and treatment of alimentary and respiratory diseases in dogs.

• Journal of agriculture & life science
• /
• v.46 no.2
• /
• pp.107-114
• /
• 2012

The present study evaluated the potential use of immunoglobulin prepared from egg yolk of chickens immunized with Escherichia coli K88 (IgY-Ec) in the control of E. coli K88 infection in RAW 264.7 murine macrophage. The binding activity of IgY-Ec against E. coli K88 surface protein was more specific and increased than control IgY. In infection assay of E. coli in macrophage, the specific IgY-Ec to E. coli K88 remarkably inhibited the phagocytic activity comparing to nonspecific IgY (p<0.001). In adherence assay, bacterial adhesion on macrophage cells was definitely reduced by preincubation of IgY-Ec compared with nonspecific IgY (p<0.05). These findings suggested that IgY-Ec have the protective effects against pathogens and IgY-based diets may have potential benefits for preventing or treating various infections in domestic animals.

• Korean Journal of Veterinary Research
• /
• v.36 no.4
• /
• pp.895-902
• /
• 1996

산란계에 불활화 개 파보바이러스 백신을 근육내로 1주 간격으로 4회 접종하여 면역화시키고 최종 접종 2주후에 채란하여 $4^{\circ}C$에 보관하며 사용하였다. 난황항체는 5% HPMCP를 이용하여 분리하였고 0.5% HPMCP 용액은 lipid 침전에 매우 효과적이었으며 희석배수 10배에서 투명한 상층액을 나타내었다. 1차분리한 상층액의 단백질 농도는 2.5mg/ml이었고 최종 단백질 용액의 경우는 26.53mg/ml이었다. SDS-PAGE 전기영동상에서 분자량 60~70 KD 및 30~40 KD의 2 band가 나타났으며 non-reducing 전기영동에서는 닭 혈청 IgG와 같은 120~160 KD의 분자량을 보인 band가 각각의 분리용액에서 나타났다. 난황 항체의 개 파보바이러스에 대한 혈구응집억제반응 항체역가는 혈청의 역가에 비해 1주의 차이를 주며 증가했으며 난황 항체는 1:640에서 1:2560, 혈청은 1:640에서 1:5120을 나타내었다.

• Korean Journal of Veterinary Research
• /
• v.36 no.4
• /
• pp.903-913
• /
• 1996

총 15두의 초유를 섭취하지 않은 신생자견을 대상으로 난황항체를 경구투여한 후 개 파보바이러스를 경구 접종하여 실험감염을 유발시켜 난황항체의 수동 면역에 의한 예방효과를 알아보고자 한다. 항체역가는 면역화된 산란계로부터 분리한 난황항체를 투여한 자견이 비면역 난황항체를 투여한 자견에 비해 높았다. 개 파보바이러스 접종 직전의 항체역가는 대조군의 경우 1:40에서 1:80, 실험군의 경우는 1:320에서 1:1280이었다. 모든 대조군의 자견들은 바이러스 접종후 4일에 임상증상을 나타내었고 총 7두중 6두가 폐사된 반면 실험군 자견은 2두만이 증상을 나타내었고 폐사 자견은 없었다(p<0.01). 개 파보바이러스를 경구 접종한 후 전체 자견의 혈구응집억제반응역가는 접종후 6일까지 감소하는 경향을 보였다. 접종후 5일의 분변내 혈구 응집반응역가는 실험군 자견의 경우 < 2에서 64였으며 대조자견은 216에서 2048로 높았다.