Muscovy duck adenovirus type-2 and type-3 refined egg yolk antibody and preparation method thereof

Abstract

The invention discloses a muscovy duck adenovirus type-2 and type-3 refined egg yolk antibody and a preparation method thereof. The method is as follows: immunizing healthy laying hens by using a duckadenovirus type-2 and type-3 combined inactivated vaccine, and collecting high-immune eggs when the titer of a muscovy duck adenovirus type-2 and type-3 agar diffusion antibody in the egg yolk of thelaying hens is greater than or equal to 1:32; separating egg yolk from the high-immune eggs and carrying out first inactivation; adding an egg yolk liquid subjected to the first inactivation into anacidification reaction tank for acidification and extraction, centrifugally separating an acidification extracting solution, adding octanoic acid into supernatant for second inactivation, performing deep filtration, and adding a formaldehyde solution into a filtrate for third inactivation; and performing concentration adjustment, pH adjustment, sterilization and filtration to obtain the muscovy duck adenovirus type-2 and type-3 refined egg yolk antibody. After the muscovy duck adenovirus type 2 and type 3 refined egg yolk antibody prepared by the method are used for immunizing muscovy ducks, the muscovy duck adenovirus type-2 and type-3 challenge test proves that the refined egg yolk antibody can effectively and better protect duckling adenoviruses type-2 and type-3.

Description

technical field

[0001] The invention relates to the technical field of veterinary biological products, in particular to a refined egg yolk antibody of muscovy duck adenovirus type 2 and type 3 and a preparation method thereof. Background technique

[0002] Avian adenovirus is one of the common infectious disease pathogens in poultry, which can cause clinical and pathological syndromes in chickens, ducks, geese and other poultry or wild birds, including hydropericardium, hepatitis, aplastic anemia, hemorrhage, A variety of poultry conditions such as mild respiratory disease.

[0003] Duck (Anas platyrhynchos domestica)-derived adenoviruses include duck-derived adenovirus type A (Duck atadenovirus A, DAdV-A) of the AT-rich adenovirus genus and duck-derived adenovirus type B (DAdV-B) of the avian adenovirus genus. DAdV-A is duck adenovirus type 1 (Duck adenovirus type 1, DAdV-1), also known as egg drop syndrome virus (Egg drop syndrome virus, EDSV), which is related to egg dro...

Examples

Embodiment 1

[0034] 1 Production of high-free eggs

[0035] 1.1 Laying hens should have the production performance of commercial laying hens.

[0036] 1.1.1 No avian leukosis and avian influenza infection According to 0.5% of the chicken flock blood sampling, all should be negative.

[0037] 1.1.2 Pullorum and Mycoplasma pullorum and Mycoplasma pullorum are detected according to NY / T536-2002 "Diagnostic Techniques for Typhoid and Pullorum" and NY / T553-2002 "Diagnostic Techniques for Mycoplasma Diseases", and the positive rate of pullorum and Mycoplasma infection should be ≤0.1 %.

[0038] 1.1.3 Feeding and management of laying hens The construction of chicken farms must meet the requirements of veterinary hygiene and epidemic prevention regulations. The chicken farm should be more than 500 meters away from the main traffic road, and the entrance and exit roads should be separated. The material and manure roads in the site are separated. There should be a disinfection pool at the entran...

Embodiment 2

[0047] Example 2: Preparation of Muscovy Duck Adenovirus Type 2 and Type 3 Refined Egg Yolk Antibodies

[0048] 2.1 Eggshell disinfection

[0049] Dip high-free eggs into 0.1% bromogeramine aqueous solution at 42°C for 15 minutes for disinfection. If the eggshells are seriously polluted, select them in advance, disinfect them separately and rinse them with clean water, then soak them again for disinfection. Subsequently, sterilize the eggs by immersing them in a water bath above 95°C for 5 seconds, take them out and dry them in the air or blow dry them for later use.

[0050] 2.2 Separation of egg yolks Beat the eggs manually or mechanically, fully remove the egg white (white), blastodisc and frenulum, and collect the egg yolks.

[0051] 2.3 Inactivation of I

[0052] Add the egg yolk liquid into a pasteurization tank, add an appropriate amount of water for injection, stir and mix evenly, heat to 65°C, keep warm for 15 minutes to inactivate, after the inactivation is complet...

Embodiment 3

[0062] Embodiment 3: Antibody test

[0063] 3.1 The purity is tested according to the current "Veterinary Pharmacopoeia of the People's Republic of China", and there is no contamination by bacteria, mycoplasma and foreign viruses.

[0064] 3.2 Caprylic acid and formaldehyde residues were tested according to the current Veterinary Pharmacopoeia of the People's Republic of China, and the results met the requirements.

[0065] 3.3 Safety test Ten 5-day-old healthy muscovy ducks were subcutaneously injected with 1.0ml, observed for 14 days, and all were healthy and alive.

[0066] 3.4 Effectiveness test

[0067] 3.4.1 Detection of antibody agar expansion titer According to the current "People's Republic of China Veterinary Pharmacopoeia", the antibody agar expansion antibody titer is tested, and the antibody result is recorded.

[0068] 3.4.2 Immune challenge method Use 40 young Muscovy ducks aged 1 to 3 days and divide them into 5 groups randomly, 10 ducks in each group. Group...

technical field

[0001] The invention relates to the technical field of veterinary biological products, in particular to a refined egg yolk antibody of muscovy duck adenovirus type 2 and type 3 and a preparation method thereof. Background technique

[0002] Avian adenovirus is one of the common infectious disease pathogens in poultry, which can cause clinical and pathological syndromes in chickens, ducks, geese and other poultry or wild birds, including hydropericardium, hepatitis, aplastic anemia, hemorrhage, A variety of poultry conditions such as mild respiratory disease.

[0003] Duck (Anas platyrhynchos domestica)-derived adenoviruses include duck-derived adenovirus type A (Duck atadenovirus A, DAdV-A) of the AT-rich adenovirus genus and duck-derived adenovirus type B (DAdV-B) of the avian adenovirus genus. DAdV-A is duck adenovirus type 1 (Duck adenovirus type 1, DAdV-1), also known as egg drop syndrome virus (Egg drop syndrome virus, EDSV), which is related to egg dro...

Examples

Embodiment 1

[0034] 1 Production of high-free eggs

[0035] 1.1 Laying hens should have the production performance of commercial laying hens.

[0036] 1.1.1 No avian leukosis and avian influenza infection According to 0.5% of the chicken flock blood sampling, all should be negative.

[0037] 1.1.2 Pullorum and Mycoplasma pullorum and Mycoplasma pullorum are detected according to NY / T536-2002 "Diagnostic Techniques for Typhoid and Pullorum" and NY / T553-2002 "Diagnostic Techniques for Mycoplasma Diseases", and the positive rate of pullorum and Mycoplasma infection should be ≤0.1 %.

[0038] 1.1.3 Feeding and management of laying hens The construction of chicken farms must meet the requirements of veterinary hygiene and epidemic prevention regulations. The chicken farm should be more than 500 meters away from the main traffic road, and the entrance and exit roads should be separated. The material and manure roads in the site are separated. There should be a disinfection pool at the entran...

Embodiment 2

[0047] Example 2: Preparation of Muscovy Duck Adenovirus Type 2 and Type 3 Refined Egg Yolk Antibodies

[0048] 2.1 Eggshell disinfection

[0049] Dip high-free eggs into 0.1% bromogeramine aqueous solution at 42°C for 15 minutes for disinfection. If the eggshells are seriously polluted, select them in advance, disinfect them separately and rinse them with clean water, then soak them again for disinfection. Subsequently, sterilize the eggs by immersing them in a water bath above 95°C for 5 seconds, take them out and dry them in the air or blow dry them for later use.

[0050] 2.2 Separation of egg yolks Beat the eggs manually or mechanically, fully remove the egg white (white), blastodisc and frenulum, and collect the egg yolks.

[0051] 2.3 Inactivation of I

[0052] Add the egg yolk liquid into a pasteurization tank, add an appropriate amount of water for injection, stir and mix evenly, heat to 65°C, keep warm for 15 minutes to inactivate, after the inactivation is complet...

Embodiment 3

[0062] Embodiment 3: Antibody test

[0063] 3.1 The purity is tested according to the current "Veterinary Pharmacopoeia of the People's Republic of China", and there is no contamination by bacteria, mycoplasma and foreign viruses.

[0064] 3.2 Caprylic acid and formaldehyde residues were tested according to the current Veterinary Pharmacopoeia of the People's Republic of China, and the results met the requirements.

[0065] 3.3 Safety test Ten 5-day-old healthy muscovy ducks were subcutaneously injected with 1.0ml, observed for 14 days, and all were healthy and alive.

[0066] 3.4 Effectiveness test

[0067] 3.4.1 Detection of antibody agar expansion titer According to the current "People's Republic of China Veterinary Pharmacopoeia", the antibody agar expansion antibody titer is tested, and the antibody result is recorded.

[0068] 3.4.2 Immune challenge method Use 40 young Muscovy ducks aged 1 to 3 days and divide them into 5 groups randomly, 10 ducks in each group. Group...

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