CN113396864A - Method for purifying leukemia of chickens - Google Patents

Method for purifying leukemia of chickens Download PDF

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CN113396864A
CN113396864A CN202110673690.7A CN202110673690A CN113396864A CN 113396864 A CN113396864 A CN 113396864A CN 202110673690 A CN202110673690 A CN 202110673690A CN 113396864 A CN113396864 A CN 113396864A
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chickens
leukemia
chicken
purifying
breeding
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王璇
文正常
张杨子
张晓可
潘淑惠
吴玙彤
徐景峨
冉江
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Guizhou Institute Of Animal Husbandry And Veterinary Medicine
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/02Breeding vertebrates
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/10Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/163Sugars; Polysaccharides
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/174Vitamins
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/70Feeding-stuffs specially adapted for particular animals for birds
    • A23K50/75Feeding-stuffs specially adapted for particular animals for birds for poultry
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983Viruses
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/08RNA viruses
    • G01N2333/15Retroviridae, e.g. bovine leukaemia virus, feline leukaemia virus, feline leukaemia virus, human T-cell leukaemia-lymphoma virus
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2469/00Immunoassays for the detection of microorganisms
    • G01N2469/10Detection of antigens from microorganism in sample from host

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Abstract

The invention belongs to the technical field of poultry breeding, and discloses a method for purifying leukemia of chickens, which comprises the following steps: establishing a safety infrastructure, and implementing safety management in the cultivation process; in the safe cultivation process, the chickens are subjected to leukemia purification; in the process of purifying the leukemia of the chickens, different detection indexes are collected according to different breeding stages, and the chickens with positive display are removed; cleaning and disinfecting the infrastructure on time in the cultivation process, placing the negative chickens into the infrastructure for expanding the seeds, and injecting vaccines into the negative chickens. Placing negative chickens into a facility for expanding seeds, taking the negative chickens as breeding seeds for isolated breeding, and placing the negative chickens in the same cage; and a blocking measure is adopted between every two cages, so that horizontal transmission is avoided, and direct contact between the cages and convection of direct air flow are avoided. Meanwhile, the invention solves the problem that the difficulty is increased in sequence by dividing the purification of the avian leukosis into three stages of reduction, control and eradication in the prior art.

Description

Method for purifying leukemia of chickens
Technical Field
The invention belongs to the technical field of poultry breeding, and particularly relates to a method for purifying leukemia of chickens.
Background
Poultry is currently referred to as artificially housed avian animals, and has other uses, primarily for the purpose of obtaining their meat, eggs and feathers. Typically animals of the Phasianidae and Anatidae families, such as chickens, ducks, geese, etc., and birds of other families, such as turkeys, pigeons, quails and various birds with a whizzy bird. Some of the land birds such as domestic chicken, turkey, guinea fowl and pigeon, and waterfowl such as duck and goose are usually raised for appreciation, while the production of eggs, meat, feathers, etc. is usually the purpose. Wherein the chicken poultry is one of poultry. In the process of breeding the chickens, leukemia is easy to appear.
Avian leukemia is a general term for a variety of neoplastic diseases caused by Avian Leukemia Virus (ALV) and Avian Sarcoma Virus (ASV). In nature, only chickens can infect avian leukemias. The resistance of chickens of different breeds or strains to viral infection and tumorigenesis varies greatly. The susceptibility of the chickens is higher than that of the cocks, the chickens mainly occur more than 18 weeks old, and the death rate is 5-6%. ALVs can spread vertically and horizontally by egg laying. Therefore, ALV has been an important disease in layer chicken breeding and egg laying.
However, the prior art divides the purification of the avian leukosis into three stages of reduction, control and eradication, which increases the difficulty in turn; meanwhile, the existing chicken leukemia purification technology has the problems of high cost and low efficiency, and the aim of completely purifying the avian leukemia cannot be achieved. Therefore, a new method for purifying the leukemia of the chicken is needed to overcome the problems and defects in the prior art.
Through the above analysis, the problems and defects of the prior art are as follows: the prior art divides the purification of the avian leukosis into three stages of reduction, control and eradication, which increases the difficulty in turn; meanwhile, the existing chicken leukemia purification technology has the problems of high cost and low efficiency, and the aim of completely purifying the avian leukemia cannot be achieved.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a method for purifying the leukemia of the chickens.
The invention is realized in such a way, and the method for purifying the leukemia of the chicken comprises the following steps:
step one, establishing a safety infrastructure, and implementing safety management in the cultivation process; the safety infrastructure comprises a station enzyme labeling instrument, a station plate washing machine, a carbon dioxide incubator, a biological safety cabinet, a station cell special microscope, a large-flux refrigerated centrifuge, an ultra-pure water machine, a drying box, a constant-temperature incubator and a pipettor; the safety management comprises the safety management of an incubation process stage, the safety management of a production process stage, the safety management of an immunization stage and the improvement of feeding management;
step two, in the safe cultivation process, the chickens are subjected to leukemia purification; in the process of purifying the leukemia of the chickens, different detection indexes are collected according to different breeding stages, and the chickens with positive display are removed; wherein, according to different stages of breeding, gather different detection index, get rid of and show positive chicken, include:
marking poultry chickens one by one at the age of 1-5 weeks, and collecting meconium; performing liquid nitrogen freeze thawing on the meconium sample, then performing ALV P27 antigen detection, and if positive detection is detected, eliminating chicks; the chick with no positive detection is fed in a group isolation way;
collecting and carrying out ALVP27 antigen detection one by one at the age of 6-10 weeks, and eliminating all positive chickens;
detecting 3 eggs laid at the beginning of 23-25 weeks old one by one, collecting semen samples one by the cocks, carrying out ALV P27 antigen detection on the egg samples and the semen, and eliminating all positive chickens;
performing spot inspection on antibodies of the AB subgroup and the J subgroup of the avian leukemia at the age of 24-25 weeks;
at the age of 35-45 weeks, detecting 3 egg whites one by one, collecting semen samples one by one for cocks, carrying out ALV P27 antigen detection on the egg samples and the semen, and eliminating all positive chickens;
after 45 weeks of age, egg white is detected one by one every 10 days until seeds are reserved, the judgment value of the embryo manure and egg white elimination standard is that S/P is more than or equal to 0.1, and the judgment value of the virus separation elimination standard is that S/P is more than or equal to 0.08;
step three, cleaning, disinfecting, expanding and injecting vaccines on time to the infrastructure in the cultivation process: the method comprises the following steps of (1) leaving the infrastructure empty for 1-2 weeks before the chickens are placed in the infrastructure; arranging, cleaning and thoroughly disinfecting all facility equipment in the stall, and cleaning and thoroughly disinfecting the surrounding field and the road of the stall; meanwhile, three different disinfection solutions are used for disinfection on the 1 st day, the 13 th day and the 14 th day; and (3) placing the chicken with negative detection into a facility for expanding the seeds, and injecting the vaccine into the chicken with negative detection.
Further, in the first step, the safety management of the incubation process stage includes: the hatching of the chicks needs to be independent, and the hatching of single strains are realized.
Further, in the first step, the safety management of the production process stage includes:
breeding in small cages in a breeding stage and breeding in single cage in an egg laying stage;
in the breeding stage, the negative chicken flocks are detected and eliminated in time, then the negative chicken flocks are moved into a chicken house which is disinfected and meets the biological safety standard, and the sanitary and safe properties are required to be paid attention to so as to improve the breeding management;
artificial insemination must be achieved by one chicken with one insemination tube to prevent cross contamination.
Further, in step one, the safety management of the immunization phase includes:
the avian leukosis virus p27 antigen is detected by the avian leukosis antigen detection kit, the use of live vaccines is reduced by the core group, and the attenuated vaccines are replaced by genetic engineering vaccines.
Further, in the first step, the improving the feeding management comprises:
fumigating the henhouse, and spraying 1-3% hot caustic soda diluent on the ground and the walls for disinfection;
after entering a chicken house, the chicks drink warm boiled water first and then eat the chicks, and a small amount of glucose, organic acid, probiotics and vitamin C are added into drinking water;
the feed is scattered on a shallow tray or plastic cloth to ensure that each chick can eat freely, and the temperature of the henhouse is controlled to be 33-38 ℃.
Further, in the second step, the ALV P27 antigen detection is carried out on the meconium sample after freeze thawing by liquid nitrogen, and the method comprises the following steps:
(1) the hatching eggs are placed in paper bags for isolated hatching, and each chicken produces the hatching eggs which are independently used by one shelling paper bag, so that the propagation is avoided; placing the meconium in a centrifuge tube, adding a sample diluent into the centrifuge tube, and fully and uniformly oscillating;
(2) quickly putting the uniformly oscillated solution into liquid nitrogen for freezing, taking out, and putting into a water bath at thirty-seven ℃ for thawing;
(3) repeating for several times to fully release the avian leukosis virus P27 antigen, centrifuging, and collecting supernatant for detecting avian leukosis virus P27 antigen in meconium; the avian leukosis virus P27 antigen is prepared with an equivalent enzyme-linked immunosorbent assay kit.
Further, the sample diluent contains ten percent by volume of PBST solution of newborn bovine serum or commercial kit-specific diluent.
Further, in the third step, the chicken birds with negative detection are put into the facility for expanding seeds, and the third step comprises the following steps:
negative chickens are used as breeding for isolated breeding, and are placed in the same cage;
and a blocking measure is adopted between every two cages, so that horizontal transmission is avoided, and meanwhile, direct contact between the cages and convection of direct air flow are avoided.
Further, in step three, the chicken poultry injection vaccine for negative detection comprises:
detecting when vaccine injection is carried out;
a vaccine produced using chick embryo derived cells as a starting material.
Further, in the third step, when the immunization is carried out on the chicken poultry injection vaccine with negative detection, the disinfection of instruments and the disinfection of hands are carried out, and simultaneously, one needle head is replaced for each chicken during the inoculation.
By combining all the technical schemes, the invention has the advantages and positive effects that: the method for purifying the avian leukosis solves the problem that the difficulty is increased in sequence by dividing the purification of the avian leukosis into three stages of reduction, control and eradication in the prior art, solves the problems of high cost and low efficiency of the existing avian leukosis purification technology, and achieves the purpose of completely purifying the avian leukosis. The invention can avoid the spreading of virus level from pancreas infected with embryo to newly hatched chicken, and avoid the problem of increasing miss rate caused by the reduction of meconium freezing and thawing times; meanwhile, the invention is used for purifying the avian leukosis for a long time, the positive rate of each avian leukosis is less than 1 percent, and the effect is obvious.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed to be used in the embodiments of the present application will be briefly described below, and it is obvious that the drawings described below are only some embodiments of the present application, and it is obvious for those skilled in the art that other drawings can be obtained from the drawings without creative efforts.
FIG. 1 is a flow chart of a method for purifying avian leukosis provided in an embodiment of the present invention.
Fig. 2 is a flowchart of a safety management method for a production process stage according to an embodiment of the present invention.
Fig. 3 is a flowchart of an improved feeding management method according to an embodiment of the present invention.
FIG. 4 is a flow chart of a method for detecting ALV P27 antigen after freezing and thawing a meconium sample by liquid nitrogen, which is provided by the embodiment of the invention.
Fig. 5 is a flowchart of a method for timely cleaning and disinfecting the infrastructure during the cultivation process according to an embodiment of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Aiming at the problems in the prior art, the invention provides a method for purifying avian leukosis, and the invention is described in detail below with reference to the accompanying drawings.
As shown in fig. 1, the method for purifying leukemia of chicken provided by the embodiment of the invention comprises the following steps:
s101, establishing a safety infrastructure, and implementing safety management in the cultivation process;
s102, in the safe cultivation process, leukemia purification is carried out on the chickens; in the process of purifying the leukemia of the chickens, different detection indexes are collected according to different breeding stages, and the chickens with positive display are removed;
s103, cleaning and disinfecting the infrastructure on time in the cultivation process, putting the negative detection chickens into the infrastructure for expanding the seeds, and injecting vaccines into the negative detection chickens.
In step S101 provided in the embodiment of the present invention, the safety infrastructure includes a stage microplate reader, a stage plate washing machine, a carbon dioxide incubator, a biosafety cabinet, a stage cell special microscope, a large-flux refrigerated centrifuge, an ultra-pure water machine, a drying oven, a constant temperature incubator, and a pipette.
In step S101 provided by the embodiment of the present invention, the safety management includes safety management in an incubation process stage, safety management in a production process stage, safety management in an immunization stage, and improvement of feeding management.
The safety management of the incubation process stage provided by the embodiment of the invention comprises the following steps: the hatching of the chicks needs to be independent, and the hatching of single strains are realized.
As shown in fig. 2, the safety management of the production process stage provided by the embodiment of the present invention includes:
s201, small-cage breeding in a breeding stage and single-cage breeding in an egg laying stage;
s202, timely detecting and eliminating are needed in the breeding stage, then negative chicken flocks are moved into a chicken house which is disinfected and meets the biological safety standard, and the sanitary and safe need to be paid attention to so as to improve the breeding management;
s203, artificial insemination must be achieved by one chicken with one insemination tube to prevent cross contamination.
The safety management of the immune stage provided by the embodiment of the invention comprises the following steps: the avian leukosis virus p27 antigen is detected by the avian leukosis antigen detection kit, the use of live vaccines is reduced by the core group, and the attenuated vaccines are replaced by genetic engineering vaccines.
As shown in fig. 3, the improved feeding management provided by the embodiment of the present invention includes:
s301, fumigating the henhouse, and spraying 1-3% hot caustic soda diluent on the ground and the walls for disinfection;
s302, after the chicks enter the henhouse, drinking warm boiled water first and then eating, and adding a small amount of glucose, organic acid, probiotics and vitamin C into drinking water;
s303, scattering the feed on a shallow tray or plastic cloth to ensure that each chick can feed freely, and controlling the temperature of the henhouse to be 33-38 ℃.
The improved feeding management provided by the embodiment of the invention further comprises the following steps: feeding organic acid and probiotics to reduce intestinal reaction, improve immunity and reduce stress of chicken flocks; the chicken coops, the feed troughs and the waterlines are made to be sanitary, and the mosquitoes, the flies and the rats are killed.
In step S102 provided in the embodiment of the present invention, the collecting different detection indexes according to different breeding stages to remove the chicken showing positive results includes:
marking poultry chickens one by one at the age of 1-5 weeks, and collecting meconium; performing liquid nitrogen freeze thawing on the meconium sample, then performing ALV P27 antigen detection, and if positive detection is detected, eliminating chicks; the chick with no positive detection is fed in a group isolation way;
collecting and carrying out ALVP27 antigen detection one by one at the age of 6-10 weeks, and eliminating all positive chickens;
detecting 3 eggs laid at the beginning of 23-25 weeks old one by one, collecting semen samples one by the cocks, carrying out ALV P27 antigen detection on the egg samples and the semen, and eliminating all positive chickens;
performing spot inspection on antibodies of the AB subgroup and the J subgroup of the avian leukemia at the age of 24-25 weeks;
at the age of 35-45 weeks, detecting 3 egg whites one by one, collecting semen samples one by one for cocks, carrying out ALV P27 antigen detection on the egg samples and the semen, and eliminating all positive chickens;
after 45 weeks of age, the egg white is detected one by one every 10 days until the seeds are reserved, the judgment value of the meconium and egg white elimination standard is that S/P is more than or equal to 0.1, and the judgment value of the virus separation elimination standard is that S/P is more than or equal to 0.08.
As shown in fig. 4, in step S102 provided by the embodiment of the present invention, the performing ALV P27 antigen detection on the meconium sample after freezing and thawing by liquid nitrogen includes:
s401, placing the hatching eggs in paper bags for isolated hatching, wherein each hatching egg laid by each chicken is independently used by one shelling paper bag, so that propagation is avoided; placing the meconium in a centrifuge tube, adding a sample diluent into the centrifuge tube, and fully and uniformly oscillating;
s402, quickly putting the uniformly oscillated solution into liquid nitrogen for freezing, taking out, and putting into a water bath at thirty-seven ℃ for thawing;
s403, repeating for several times to fully release the avian leukosis virus P27 antigen, centrifuging and taking supernatant to detect the avian leukosis virus P27 antigen in meconium; the avian leukosis virus P27 antigen is prepared with an equivalent enzyme-linked immunosorbent assay kit.
The sample diluent provided by the embodiment of the invention contains ten percent of PBST solution of newborn bovine serum or special diluent of a commercial kit.
As shown in fig. 5, in step S103, the timely cleaning and disinfecting the infrastructure during the cultivation process includes:
s501, before the chickens are placed in the infrastructure, the infrastructure is empty for 1-2 weeks;
s502, arranging, cleaning and thoroughly disinfecting all facility equipment in the stall, and cleaning and thoroughly disinfecting the surrounding field and the road of the stall;
s503, sterilizing by using three different sterilizing solutions on the 1 st day, the 13 th day and the 14 th day; and (3) placing the chicken with negative detection into a facility for expanding the seeds, and injecting the vaccine into the chicken with negative detection.
In step S103 provided by the embodiment of the present invention, the placing the negative detection chickens into a facility for expanding the breeds includes:
negative chickens are used as breeding for isolated breeding, and are placed in the same cage;
and a blocking measure is adopted between every two cages, so that horizontal transmission is avoided, and meanwhile, direct contact between the cages and convection of direct air flow are avoided.
In step S103 provided by the embodiment of the present invention, the step of injecting the vaccine into the detection negative chickens includes:
detecting when vaccine injection is carried out;
a vaccine produced using chick embryo derived cells as a starting material.
In step S103 provided by the embodiment of the present invention, when the negative chicken vaccine is immunized, the device and the hands are sterilized, and a needle is replaced for each chicken during the inoculation.
In the description of the present invention, "a plurality" means two or more unless otherwise specified; the terms "upper", "lower", "left", "right", "inner", "outer", "front", "rear", "head", "tail", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, are only for convenience in describing and simplifying the description, and do not indicate or imply that the device or element referred to must have a particular orientation, be constructed in a particular orientation, and be operated, and thus, should not be construed as limiting the invention. Furthermore, the terms "first," "second," "third," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention, and the scope of the present invention is not limited thereto, and any modification, equivalent replacement, and improvement made by those skilled in the art within the technical scope of the present invention disclosed herein, which is within the spirit and principle of the present invention, should be covered by the present invention.

Claims (10)

1. The method for purifying the leukemia of the chickens is characterized by comprising the following steps of:
step one, establishing a safety infrastructure, and implementing safety management in the cultivation process; the safety infrastructure comprises a station enzyme labeling instrument, a station plate washing machine, a carbon dioxide incubator, a biological safety cabinet, a station cell special microscope, a large-flux refrigerated centrifuge, an ultra-pure water machine, a drying box, a constant-temperature incubator and a pipettor; the safety management comprises the safety management of an incubation process stage, the safety management of a production process stage, the safety management of an immunization stage and the improvement of feeding management;
step two, in the safe cultivation process, the chickens are subjected to leukemia purification; in the process of purifying the leukemia of the chickens, different detection indexes are collected according to different breeding stages, and the chickens with positive display are removed; wherein, according to different stages of breeding, gather different detection index, get rid of and show positive chicken, include:
marking poultry chickens one by one at the age of 1-5 weeks, and collecting meconium; performing liquid nitrogen freeze thawing on the meconium sample, performing ALVP27 antigen detection, and if positive detection is detected, eliminating chicks; the chick with no positive detection is fed in a group isolation way;
collecting and carrying out ALVP27 antigen detection one by one at the age of 6-10 weeks, and eliminating all positive chickens;
detecting 3 eggs laid at the beginning of 23-25 weeks old one by one, collecting semen samples one by the cocks, carrying out ALVP27 antigen detection on the egg samples and the semen, and eliminating all positive chickens;
performing spot inspection on antibodies of the AB subgroup and the J subgroup of the avian leukemia at the age of 24-25 weeks;
at the age of 35-45 weeks, detecting 3 egg whites one by one, collecting semen samples one by one for cocks, and carrying out ALVP27 antigen detection on the egg samples and the semen to eliminate all positive chickens;
after 45 weeks of age, egg white is detected one by one every 10 days until seeds are reserved, the judgment value of the embryo manure and egg white elimination standard is that S/P is more than or equal to 0.1, and the judgment value of the virus separation elimination standard is that S/P is more than or equal to 0.08;
step three, cleaning, disinfecting, expanding and injecting vaccines on time to the infrastructure in the cultivation process: the method comprises the following steps of (1) leaving the infrastructure empty for 1-2 weeks before the chickens are placed in the infrastructure; arranging, cleaning and thoroughly disinfecting all facility equipment in the stall, and cleaning and thoroughly disinfecting the surrounding field and the road of the stall; meanwhile, three different disinfection solutions are used for disinfection on the 1 st day, the 13 th day and the 14 th day; and (3) placing the chicken with negative detection into a facility for expanding the seeds, and injecting the vaccine into the chicken with negative detection.
2. The method for purifying leukemia in poultry according to claim 1, wherein in the first step, the safety management of the hatching process stage comprises: the hatching of the chicks needs to be independent, and the hatching of single strains are realized.
3. The method for purifying leukemia of chicken according to claim 1, wherein the safety management of the production process stage in the first step comprises:
breeding in small cages in a breeding stage and breeding in single cage in an egg laying stage;
in the breeding stage, the negative chicken flocks are detected and eliminated in time, then the negative chicken flocks are moved into a chicken house which is disinfected and meets the biological safety standard, and the sanitary and safe properties are required to be paid attention to so as to improve the breeding management;
artificial insemination must be achieved by one chicken with one insemination tube to prevent cross contamination.
4. The method for purifying leukemia in poultry according to claim 1, wherein the safety management of the immunization stage in the first step comprises:
the avian leukosis virus p27 antigen is detected by the avian leukosis antigen detection kit, the use of live vaccines is reduced by the core group, and the attenuated vaccines are replaced by genetic engineering vaccines.
5. The method for purifying leukemia in poultry according to claim 1, wherein the improvement of the feeding management in the first step comprises:
fumigating the henhouse, and spraying 1-3% hot caustic soda diluent on the ground and the walls for disinfection;
after entering a chicken house, the chicks drink warm boiled water first and then eat the chicks, and a small amount of glucose, organic acid, probiotics and vitamin C are added into drinking water;
the feed is scattered on a shallow tray or plastic cloth to ensure that each chick can eat freely, and the temperature of the henhouse is controlled to be 33-38 ℃.
6. The method for purifying leukemia of chicken as claimed in claim 1, wherein in the second step, the ALVP27 antigen detection is carried out after the meconium sample is frozen and thawed by liquid nitrogen, which comprises the following steps:
(1) the hatching eggs are placed in paper bags for isolated hatching, and each chicken produces the hatching eggs which are independently used by one shelling paper bag, so that the propagation is avoided; placing the meconium in a centrifuge tube, adding a sample diluent into the centrifuge tube, and fully and uniformly oscillating;
(2) quickly putting the uniformly oscillated solution into liquid nitrogen for freezing, taking out, and putting into a water bath at thirty-seven ℃ for thawing;
(3) repeating for several times to fully release the avian leukosis virus P27 antigen, centrifuging, and collecting supernatant for detecting avian leukosis virus P27 antigen in meconium; the avian leukosis virus P27 antigen is prepared with an equivalent enzyme-linked immunosorbent assay kit.
7. The method for purifying leukemia of chicken according to claim 6, wherein the sample diluent comprises ten percent by volume of PBST solution of newborn calf serum or diluent specially used for commercial kits.
8. The method for purifying leukemia of poultry according to claim 1, wherein in step three, the poultry with negative detection is put into a facility for expansion, comprising:
negative chickens are used as breeding for isolated breeding, and are placed in the same cage;
and a blocking measure is adopted between every two cages, so that horizontal transmission is avoided, and meanwhile, direct contact between the cages and convection of direct air flow are avoided.
9. The method for purifying avian leukosis of claim 1, wherein the step three, the step of injecting the vaccine into the test-negative avian comprises:
detecting when vaccine injection is carried out;
a vaccine produced using chick embryo derived cells as a starting material.
10. The method for purifying leukemia of poultry and fowl according to claim 1, wherein in the third step, when immunization is performed on the chicken and fowl injection vaccine which is negative in detection, instrument disinfection and hand disinfection are performed, and a needle is replaced for each chicken and fowl at the same time of inoculation.
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