CN114532295B - Combined purification method for leukemia and pullorum disease of high-quality chicken breeding core group - Google Patents
Combined purification method for leukemia and pullorum disease of high-quality chicken breeding core group Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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Abstract
The invention discloses a method for jointly purifying leukemia and pullorum disease of a high-quality chicken breeding core group, which is characterized in that through the establishment of a biological safety system of a breeding purification plant, the method is strictly regulated for production matched management, and is based on the perfect biological safety system of the breeding purification plant and the standard production matched management, the method is mainly combined with the breeding production process to synchronously carry out the combined purification of the leukemia and pullorum disease in the same chicken group, and through the multi-item detection of fetal manure, blood plasma, semen and egg white, the separation and judgment standard of the virus of the leukemia of the poultry is improved to S/P not less than 0.08, and the detection accuracy is improved. The method has the advantages that the purification effect is good, the positive rate of the fowl leukemia in the breeding core group of high-quality chicken breeds is less than 0.2%, the positive rate of the fowl white diarrhea is less than 0.5%, the effect is remarkable, and the purification problem of fowl leukemia and fowl white diarrhea is solved from the source.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a method for combined purification of leukemia and pullorum disease in a high-quality chicken breeding core group.
Background
Leukemia and pullorum disease of birds are important diseases that jeopardize the poultry industry, causing a great deal of economic loss each year. Avian leukemia (Avian leukosis, AL) is an Avian neoplastic disease caused by Avian Leukemia Virus (ALV) of retrovirus family, in recent years, avian leukemia is widely prevalent in yellow feather chicken breeds in China, the incidence rate rises year by year, and related researches show that chicken flock infection ALV not only causes direct morbidity and death caused by tumors, but also seriously affects the production performance of chickens, delay sexual maturity, shortened egg laying peak maintenance time, reduced laying rate and reduced fertility rate, and especially can seriously affect the hatching rate of hatching eggs and the quality of offspring chicken flocks through vertical infection; salmonella pullorum is a common infectious disease, and after the breeding hens are infected, the hatching rate of hatching eggs and the survival rate of chickens can be seriously influenced. The fowl leukemia and the pullorum disease salmonella have the characteristics of horizontal transmission and vertical transmission through eggs, so far, no effective vaccine and medicine for preventing and treating fowl leukemia are available, the pullorum disease is difficult to control and easy to repeat in the production process of breeding hens, and the transmission of the two diseases can be effectively controlled only by population purification and elimination of positive chickens. In the prior art, the fowl leukemia and pullorum disease are independently purified, a scientific and reasonable program is lacked, the sampling times are high, the workload is large, the stress of chicken flocks is large, the purification cost is high, and the comprehensive purification effect is poor. Comparative document 1, application number: 202111360869.3, name of invention: a purification method of a daylily-spicy chicken leukemia comprises the following steps: uniformly arranging specific light sources in the henhouse, and setting specific feeding density; feeding vitamins regularly; selecting target purified breeding chicken groups, eliminating positive chickens to construct F0 generation breeding populations, hatching eggs to generate F1 generation breeding populations, respectively carrying out antigen detection at different time to eliminate positive chickens, height, stature and height detection and screening, constructing F1 generation breeding purified groups, hatching eggs to generate F2 generation breeding populations; and by analogy, F4-F6 generation breeding purification groups are obtained, and the purification groups are obtained when 300-day-old antigen detection reaches zero detection rate. The comparison document is different from the method of the application.
Disclosure of Invention
The invention aims to provide a method for simultaneously purifying avian leukosis and pullorum disease in the same chicken flock by perfecting a biological safety system of a breeding purification plant and carrying out standard production matched management and combining a breeding production process, and the method improves the avian leukosis virus separation judgment standard to S/P (S/P) of more than or equal to 0.08 and improves the detection accuracy by detecting multiple items of meconium, blood plasma, semen and egg white, thereby solving the purification problem of avian leukemia and pullorum disease from the source.
The invention is realized by the following technical scheme: a method for purifying leukemia and pullorum disease in high-quality chicken breeding core group is characterized in that the purification of leukemia and pullorum disease is achieved from the source through establishment of a biological safety system of a breeding purification plant, standard and strict production and matched management, a combined purification program of leukemia and pullorum disease, and through multi-science field project detection of fowl diseases, the separation judgment detection rate of leukemia is improved.
The purifying method comprises the following steps:
(1) Biological safety system establishment of breeding purification plant
A laboratory infrastructure matching with the purification requirements;
the field layout is scientific and reasonable, the living area, the production area, the sewage treatment area and the innocent treatment area of the dead chicken are separated by enclosing walls or guardrails, the distance between the areas is not less than 50 meters, the brood shed, the breeding shed, the hatching shed and the isolation shed are respectively arranged in different areas, and the mutual distance between the henhouses is not less than 15 meters;
c, the construction hardware of the field is complete, the production sewage is collected and treated in a centralized way, a vehicle disinfection pool and a personnel disinfection channel are arranged at the inlet of the life management area, the personnel disinfection channel and a shower room are arranged at the inlet of the production area, greening is not planted in the purification area, and hardening treatment is adopted on the whole field road;
(2) Standard strict production matching management
And A, hatching and matching: setting an independent breeding purification hatching hall, an independent single incubator and a hatching machine, and adopting single strain hatching and single family hatching;
b chicken house is matched with: adopting brooding to cultivate into an integral house for feeding, without transferring groups during the period, and carrying out family-based feeding in brooding stage; single cage feeding in egg laying stage, installing filtering water purifying equipment, automatic drinking water equipment and automatic medicine adding equipment, and installing rat preventing, bird preventing and mosquito preventing facilities; a disinfection pond and a disinfection hand basin are arranged at the entrance of the chicken house, and an automatic spraying disinfection device is arranged in the chicken house;
c, management matching: the feed enters a material room of each house after being uniformly fumigated and disinfected in a field area; after detection and elimination in the breeding stage, carrying the negative chicken flock into an egg laying house which is washed clean and sterilized and qualified in 3 days; manually inseminating one chicken with one insemination rubber head or insemination tube;
d, immunization link: the whole process of high standard strict management of medicines and vaccine input products is enhanced, the vaccine used in a purification field is detected, and the use without avian leukosis virus pollution is detected; in the immunization instrument sterilization and hand sterilization, each chicken is replaced by a needle;
(3) Combined purification of leukemia and pullorum disease of poultry
Chicken age selection for different weeks: age 1 day: detecting leukemia of birds, detecting full group by using meconium ELISA, and eliminating positive chicks or eliminating chicks of the same family and father and mother thereof according to the number of chicken groups;
before the egg laying shed is rotated: pullorum detection, complete group-by-group serum plate agglutination detection, negative chicken turning egg laying house, and positive chicken elimination;
20 weeks of age: pullorum disease detection, complete group serum plate agglutination detection, and positive chicken elimination;
20-21 weeks of age: avian leukosis detection, whole group virus-by-virus separation ELISA detection, and elimination of positive chickens;
21 weeks of age: avian leukemia detection, cock sperm virus separation ELISA detection, and elimination of positive chickens;
22-25 weeks of age: detecting leukemia of the poultry, detecting 3 primary egg white ELISA detection of the hen one by one, and eliminating positive chickens;
35-45 weeks of age: avian leukemia and pullorum disease detection, ELISA detection for separating avian leukemia virus from whole group to individual avian leukemia virus before seed retention, and pullorum disease plasma plate agglutination detection, and positive chicken is eliminated;
35-45 weeks of age: detecting leukemia of the poultry, separating and detecting semen viruses of the roosters before reserving the breeds, and eliminating positive chickens;
35-45 weeks of age: detecting leukemia of the poultry, detecting 2 egg white ELISA detection of the hen by hen, and eliminating positive chickens;
elimination standard: the standard judgment value of the elimination of the poultry leukemia meconium is more than or equal to 0.1, and the standard judgment value of the separation and elimination of the egg white and the virus is more than or equal to 0.08; the elimination standard of pullorum disease is judged according to NY/T536-2017 diagnosis technique for pullorum disease and typhoid.
The laboratory infrastructure comprises a biosafety cabinet, a drying oven, a constant temperature incubator, a carbon dioxide incubator, a high-pressure steam sterilization pot, a pipettor, an enzyme label instrument, a plate washer, a high-throughput freezing high-speed centrifuge, a microscope special for cells, an ultra-low temperature refrigerator at-80 ℃ and an ultra-pure water machine.
The laboratory of the breeding purification plant is equipped with instruments and equipment related to cell culture, virus separation, ELISA test and plate agglutination test.
The production function area management is enhanced, the functional areas are isolated, personnel and production materials come in and go out for disinfection management, and rat-proof, bird-proof and mosquito-proof facilities are arranged.
The production link is strictly managed, the breeding hens are bred into an integral house for breeding without transferring groups, single-cage breeding is carried out in an egg laying stage, production water is purified, an acidifier is added to inhibit growth of escherichia coli, salmonella and the like, chickens are disinfected periodically, breeding hens manually inseminate and execute one chicken insemination rubber head or seminiferous duct, hatching eggs are picked up for 3 days for 3 fumigation and independently hatched, single strains are independently hatched and independently hatched, when the hatching eggs fall on a tray, the hatching eggs of individual hens need to be independently hatched in a box, vaccine used in the whole course is used, and the poultry leukemia virus pollution is detected.
Serum, plasma, egg white and semen, serum, plasma and egg white are collected from the hen in the purification, serum, plasma and semen are collected from the cock in the purification, serum or plasma is used for the flat plate agglutination test of pullorum-chicken typhoid multivalent agglutination antigen; egg white is used for ELISA detection of ALV-P27 antigen d; plasma and cock semen were used for virus isolation by inoculation of DF-1 cells and subsequent ELISA detection of ALV-P27 antigen.
The meconium and egg white detection method specifically comprises the steps of detecting whole groups one by one and detecting the whole groups multiple times, wherein the egg white and virus separation elimination standard judgment value is increased to be more than or equal to 0.08.
The pullorum antibody detection is a plate agglutination test and is detected by serum.
The combined purification procedure is to start purification from any feeding stage of the breeding core group, select the time point and the corresponding method to implement combined purification of avian leukosis and pullorum disease.
Compared with the prior art, the method has the beneficial effects that through the establishment of a biological safety system of a breeding purification plant, strict standard production matched management and combined purification of the avian leukemia and the pullorum disease, and the basis of the perfect biological safety system of the breeding purification plant and the standard production matched management, the combined purification of the avian leukemia and the pullorum disease is mainly carried out in the same chicken flock by combining with a breeding production process, and through multi-item detection of fetal manure, blood plasma, semen and egg white, the separation judgment standard of the avian leukemia virus is improved to the S/P of more than or equal to 0.08, and the detection accuracy is improved. After 3-4 generations of combined purification, the method provided by the invention is used for combined purification of the leukemia of the fowl and the white diarrhea of the fowl, the positive rate of the leukemia of the fowl in the breeding core group of local high-quality chicken varieties such as Guangxi Sanhuang chicken, guangxi Ma chicken, yao chicken, wenchang chicken and Qingyuan chicken is less than 0.2%, the positive rate of the white diarrhea of the fowl is less than 0.5%, the effect is obvious, and the purification difficulty of the leukemia of the fowl and the white diarrhea of the fowl is solved from the source.
Drawings
FIG. 1 is a technical scheme of a combined purification method for leukemia and pullorum disease in a core group of high-quality chicken breeding.
Detailed Description
The invention will be described in detail below using the drawings and examples
Example 1, the specific method steps are:
1. biological safety system establishment of breeding purification plant
(1) The breeding purification area is constructed and improved in the early purification stage:
the field A is scientific and reasonable in layout, is far away from villages and traffic channels and is far away from migratory routes of birds. The living area, the production area, the sewage treatment area and the innocent treatment area of the dead chicken are separated by enclosing walls or guardrails, the distance between the areas is not less than 50 meters, and a brooding house, a breeding house, a hatching house, an isolation house and the like are respectively arranged in different areas, and the mutual distance between the henhouses is not less than 15 meters. Strengthening partition management, isolating functional areas, sterilizing personnel and production materials, and installing rat, bird and mosquito preventing facilities.
B, the construction hardware of the field is complete, and the production sewage is collected and treated in a centralized way, so that cross infection is reduced; the entrance of the living management area is provided with a vehicle disinfection pool and a personnel disinfection channel, and the entrance of the production area is provided with a personnel disinfection channel and a shower room. The purifying area is not planted in a greening way, so that virus and epidemic disease pollution caused by the stay of birds and mice is reduced, and the whole field road is hardened.
(2) The detection center of the purification field is provided with instruments and equipment related to cell culture, virus separation, ELISA test and plate agglutination test aiming at thoroughly eradicating avian leukemia and pullorum disease. Laboratory infrastructure to meet decontamination requirements is set up at the beginning of decontamination: there are 2 biological clean safety cabinets, 1 drying oven, 2 constant temperature incubator, 2 carbon dioxide incubator, 1 high pressure steam sterilizing pot, 2 sets of pipettors, 1 enzyme label instrument, 4 plate washer, 1 high throughput frozen high speed centrifuge, 1 microscope special for cells, 1-80 deg.c ultralow temperature refrigerator, 1 ultrapure water machine.
(3) Strict production matching management
And A, hatching and matching: setting an independent purifying hatching hall, an independent single incubator and a hatching machine, and adopting single-line hatching and single-family hatching; the hatching eggs are picked up for 3 days for fumigation and disinfection, are independently hatched, and are independently hatched and independently hatched in a single strain, and when the hatching eggs fall on a tray, the hatching eggs of individual hens need to be independently hatched in a hatching box.
B chicken house is matched with: the brooding is adopted to cultivate into an integral house for feeding, so that the early-stage cross infection is reduced, and single-cage feeding is realized in the egg laying stage. Installing a filtering type water purifying device, purifying the drinking water of the breeding hens, and adding an acidulant to inhibit the growth of escherichia coli and salmonella. An automatic drinking device and an automatic dosing device are adopted. Installing a rat-proof, bird-proof and mosquito-proof facility; the chicken house is provided with a disinfection pool at the entrance and a disinfection hand basin, and an automatic spray disinfection device is arranged in the chicken house to disinfect chicken regularly.
And C, managing and matching, namely enabling the feed to enter a material room of each house after being subjected to unified fumigation and disinfection in a purifying field. After detection and elimination in the breeding stage, the negative chicken flocks are transferred into an egg laying house which is washed cleanly and sterilized and qualified after 3 days; during the beginning of the birth period, care should be taken to avoid cross contamination, and artificial insemination is performed on one chicken with one insemination rubber head or insemination tube;
d, immunization link: the method has the advantages that the whole process of high standard strict management of medicines and vaccine input products is enhanced, and the vaccine used in the whole process of a purification field is required to be detected in advance and used without avian leukosis virus pollution;
3. combined purification procedure for leukemia and pullorum disease of poultry
1 day old, detecting fowl leukemia, detecting the total group by using meconium ELISA, and eliminating positive chicks or the chicks of the same family and father and mother of the chicks according to the number of the chicken groups;
before turning the egg laying house, detecting pullorum disease, performing agglutination detection on the whole group by serum flat plates, turning the egg laying house by negative chickens, and eliminating positive chickens;
detecting pullorum disease, detecting the agglutination of the whole group by serum flat plate, eliminating positive chicken;
detecting fowl leukemia, separating ELISA detection of whole group virus by virus, eliminating positive chicken;
avian leukemia detection, cock individual semen virus separation ELISA detection, eliminating positive chicken;
22-25 weeks of age: detecting leukemia of the poultry, detecting 3 primary egg white ELISA detection of the hen one by one, and eliminating positive chickens;
detecting fowl leukemia and pullorum disease at 35-45 weeks, separating fowl leukemia virus ELISA detection, and pullorum blood plasma plate agglutination detection before seed reserving, eliminating positive chicken;
detecting leukemia of fowl at 35-45 weeks, separating semen virus of cock before seed reserving, and eliminating positive chicken;
35-45 weeks of age: and (3) detecting leukemia of the poultry, namely detecting 2 egg white ELISA (enzyme-linked immunosorbent assay) of hen by hen, and eliminating positive chickens.
Elimination standard: the standard judgment value of meconium and egg white elimination is more than or equal to 0.1, and the standard judgment value of virus separation elimination is more than or equal to 0.08; the elimination standard of pullorum disease is judged according to NY/T536-2017 diagnosis technique for pullorum disease and typhoid.
4. The sample requirement and application are that serum, plasma, egg white and semen are collected from hen in purification, and serum, plasma and semen are collected from cock in purification. Serum or plasma is used for a plate agglutination test of pullorum-chicken typhoid multivalent agglutination antigen; egg white is used for ELISA detection of ALV-P27 antigen d; plasma and cock semen were used for virus isolation by inoculation of DF-1 cells and subsequent ELISA detection of ALV-P27 antigen.
5. The purifying and detecting process shows that the whole group of meconium, egg white and virus are separated and detected one by one, and the whole process is detected for multiple times to prevent the mixed detection of missed detection; the pullorum disease detection adopts a serum flat plate agglutination test, does not need whole blood detection, and has higher accuracy and high purification efficiency; the method is carried out in the same strain of chicken group, and the method comprises the steps of detecting the eliminated chicken white diarrhea positive chicken and detecting the eliminated fowl leukemia positive chicken, so that the purification cost is reduced. The ELISA detection elimination judgment standard of the avian leukosis is improved, positive chickens and suspicious chickens are eliminated completely, and the standard is improved to ensure the purification effect.
In example 2, the breeds of Guangxi Sanhuang chickens, guangxi Ma chickens, yao chickens, wenchang chickens, qingyuan chickens and the like are taken as examples, the positive rate of the fowl leukemia of the breeding core group through the combined purification of 3-4 generations is less than 0.2%, wherein the breeding core group of Guangxi Sanhuang chickens achieves full group negativity, the positive rate of pullorum disease is less than 0.5%, the effect is remarkable, and compared with the single purification fowl leukemia or pullorum disease in the prior art, the detection of various samples of embryo manure, blood plasma, semen and egg white is adopted, the separation judgment standard of fowl leukemia virus is improved to be more than or equal to 0.08, the detection accuracy is improved, and the purification effect is good.
The above preferred embodiments are not intended to limit the present invention, and it is possible for those skilled in the art to modify the technical solutions described in the above embodiments or to make equivalent substitutions for some technical features thereof, but any modification, equivalent substitution, improvement, etc. within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (1)
1. A method for purifying leukemia and pullorum disease in high-quality chicken breeding core group is characterized in that the purification of leukemia and pullorum disease is achieved from the source through establishment of a biological safety system of a breeding purification plant, standard and strict production and matched management, a combined purification procedure of leukemia and pullorum disease, and through multi-science field project detection of fowl diseases, the separation judgment detection rate of leukemia is improved;
the purifying method comprises the following steps:
(1) Biological safety system establishment of breeding purification plant
A laboratory infrastructure matching with the purification requirements;
the field layout is scientific and reasonable, the living area, the production area, the sewage treatment area and the innocent treatment area of the dead chicken are separated by enclosing walls or guardrails, the distance between the areas is not less than 50 meters, the brood shed, the breeding shed, the hatching shed and the isolation shed are respectively arranged in different areas, and the mutual distance between the henhouses is not less than 15 meters;
c, the construction hardware of the field is complete, the production sewage is collected and treated in a centralized way, a vehicle disinfection pool and a personnel disinfection channel are arranged at the inlet of the life management area, the personnel disinfection channel and a shower room are arranged at the inlet of the production area, greening is not planted in the purification area, and hardening treatment is adopted on the whole field road;
(2) Standard strict production matching management
And A, hatching and matching: setting an independent breeding purification hatching hall, an independent single incubator and a hatching machine, and adopting single strain hatching and single family hatching;
b chicken house is matched with: adopting brooding to cultivate into an integral house for feeding, without transferring groups during the period, and carrying out family-based feeding in brooding stage; single cage feeding in egg laying stage, installing filtering water purifying equipment, automatic drinking water equipment and automatic medicine adding equipment, and installing rat preventing, bird preventing and mosquito preventing facilities; a disinfection pond and a disinfection hand basin are arranged at the entrance of the chicken house, and an automatic spraying disinfection device is arranged in the chicken house;
c, management matching: the feed enters a material room of each house after being uniformly fumigated and disinfected in a field area; after detection and elimination in the breeding stage, carrying the negative chicken flock into an egg laying house which is washed clean and sterilized and qualified in 3 days; manually inseminating one chicken with one insemination rubber head or insemination tube;
d, immunization link: the whole process of high standard strict management of medicines and vaccine input products is enhanced, the vaccine used in a purification field is detected, and the use without avian leukosis virus pollution is detected; in the immunization instrument sterilization and hand sterilization, each chicken is replaced by a needle;
(3) Combined purification of leukemia and pullorum disease of poultry
Chicken age selection for different weeks: age 1 day: detecting leukemia of birds, detecting full group by using meconium ELISA, and eliminating positive chicks or eliminating chicks of the same family and father and mother thereof according to the number of chicken groups;
before the egg laying shed is rotated: pullorum detection, complete group-by-group serum plate agglutination detection, negative chicken turning egg laying house, and positive chicken elimination;
20 weeks of age: pullorum disease detection, complete group serum plate agglutination detection, and positive chicken elimination;
20-21 weeks of age: avian leukosis detection, whole group virus-by-virus separation ELISA detection, and elimination of positive chickens;
21 weeks of age: avian leukemia detection, cock sperm virus separation ELISA detection, and elimination of positive chickens;
22-25 weeks of age: detecting leukemia of the poultry, detecting 3 primary egg white ELISA detection of the hen one by one, and eliminating positive chickens;
35-45 weeks of age: avian leukemia and pullorum disease detection, ELISA detection for separating avian leukemia virus from whole group to individual avian leukemia virus before seed retention, and pullorum disease plasma plate agglutination detection, and positive chicken is eliminated;
35-45 weeks of age: detecting leukemia of the poultry, separating and detecting semen viruses of the roosters before reserving the breeds, and eliminating positive chickens;
35-45 weeks of age: detecting leukemia of the poultry, detecting 2 egg white ELISA detection of the hen by hen, and eliminating positive chickens;
elimination standard: the standard judgment value of the elimination of the poultry leukemia meconium is more than or equal to 0.1, and the standard judgment value of the separation and elimination of the egg white and the virus is more than or equal to 0.08; the elimination standard of the pullorum disease is judged according to NY/T536-2017 diagnosis technique of chicken typhoid and pullorum disease;
the laboratory infrastructure comprises a biosafety cabinet, a drying oven, a constant temperature incubator, a carbon dioxide incubator, a high-pressure steam sterilization pot, a pipettor, an enzyme label instrument, a plate washer, a high-throughput freezing high-speed centrifuge, a microscope special for cells, an ultra-low temperature refrigerator at-80 ℃ and an ultra-pure water machine; the laboratory of the breeding purification field is provided with instruments and equipment related to cell culture, virus separation, ELISA test and plate agglutination test; the production function distinction management is enhanced, the functional areas are isolated, personnel and production materials come in and go out for disinfection management, and rat-proof, bird-proof and mosquito-proof facilities are arranged; the production link is strictly managed, the breeding hens are bred into an integral house for breeding without transferring groups, single-cage breeding is carried out in an egg producing stage, production water is purified, an acidifier is added for inhibiting the growth of escherichia coli and salmonella, the chicken house is periodically disinfected with chickens, the breeding hens manually inseminate one chicken with insemination glue head or insemination tube, the eggs are picked up for 3 days for fumigation and disinfection, and are independently hatched, a single strain is independently hatched and independently hatched, when the eggs fall on a tray, the eggs of the individual hens need to be independently hatched in a hatching box, and vaccines used in the whole process are used for detecting the pollution of the avian-free leukemia viruses; the specific method comprises collecting serum, plasma, egg white and semen from hen in purification, collecting serum, plasma and semen from cock in purification, and performing plate agglutination test of pullorum-fowl typhoid multivalent agglutination antigen; egg white is used for ELISA detection of ALV-P27 antigen d; plasma and cock semen were used for virus isolation by inoculation of DF-1 cells and subsequent ELISA detection of ALV-P27 antigen; the detection of meconium and egg white comprises detecting whole group for multiple times, wherein the judgment value of separation and elimination standard of egg white and virus is increased to S/P not less than 0.08; the pullorum antibody detection is a plate agglutination test, and serum detection is used; the combined purification procedure is to start purification from any feeding stage of breeding core group, select time point and corresponding method to implement combined purification of fowl leukemia and pullorum disease.
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