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Production of Chicken Antibodies Against Canine Parvovirus Variant CPV-2a and the Toxicological Assessment

Author: WeiShengNan
Tutor: PengGuangNeng
School: Sichuan Agricultural University
Course: Clinical Veterinary Medicine
Keywords: egg yolk antibody CPV improved diluted with water-ammoniumsulfate protein content potency security check
CLC: S852.4
Type: Master's thesis
Year: 2013
Downloads: 30
Quote: 0
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In order to product the anti-CPV-2a type canine parvovirus antibody of yolk, and test the antibodies’toxicity safety, in the experiment, select the canine faeces which were diagnosed of canine parvovirus disease, and identification of CPV-2by PCR. Then cat kidney cells were used to purified and amplified, and collected these cells after Cytopathic effect occurred. Alternate freezing and thawing the cells three times to get virus which would be used as antigen, and storage the virus at-20℃. The virus diluted to30μg/mL dose and mixed adjuvant with ratio of1:1.After emulsifying completely, immune25-week-old laying hens. Selection of vaccine strains and vaccine virus strains of CPV-2a mixed group as an experimental control group.1825-weeks-old partridge shank chicken were divided into3groups randomly, then each group immunised with CPV-2a、 CPV-2and the mixture of CPV-2a and CPV-2separately for three times. Eggs were collected and marked. The method of improved water dilution coordinate with ammonium sulfate salting out was used to purify the chicken antibodies. SDS-PAGE, UV spectrophotometry, hemagglutination inhibition test were used to detect the purity and titer of chicken egg yolk antibodies (IgY), and also drew the development curve with these data. The results show that the titer of each group is214、212、211, and the purity is91%、91%、90.7%.Antibody titers measured with statistical analysis, the difference between group1and2was significant. Difference between group1and3is the same, significant different. Titers law drawn from the curve can be informed that in a week or so after the first immunization, antibodies can be detected in the yolk. The antibody titer reached a peak of222, since the antibody titer will be stable in the range of212-218,and maintained at this level for a long period of time. One month after the last immunization, no significant decrease in the antibody titer, which also can reach at215.The yolk antibodies test in mice for acute toxicity,28-day feeding experiment, reproductive genotoxicity tests. The acute toxicity test results showed that each mouse oral LD50more than20mL/kg BW, according this data we could judge the egg yolk antibody is non-toxic. Compared the experimental groups and control group, subacute toxicity test results were all in the normal range and found antibodies have no significant chronic toxic effects. In the micronucleus test and sperm abnormality experiment, two experimental groups’results were negative, dedicate that the anti-CPV-2a yolk antibody do no harm to mammalian chromosome and reproductive cells. The experimental results showed that the antibodies are safe and non-toxic, can be further used in food additives and clinical treatmentThe method of producing anti-CPV-2a yolk antibody was be described. Purification that was improved at the traditional method has much more greatly success rate on egg yolk antibody purification. Provide a reference for the preparation of the yolk antibody against other antigens. Meanwhile, the security of the antibody was detected, laid the foundation for the further application of egg yolk antibody.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Animal Immunology
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