108 results about "Specific-pathogen-free" patented technology

The methods provided herein relate to swine production. Specifically the instant invention provides methods for the production of terminal parent animals (i.e. terminal sires and terminal dams) for use in swine production herds. Also provided are methods for quickly and efficiently introducing and/or fixing one or more desirable traits or alleles in a swine herd. Alternatively, the present methods may be employed to eliminate a particular undesirable trait or gene. The invention also provides for herds that have been developed using any of the methods described herein. The invention also provides for the use of embryo transfer, including markerassisted embryo transfer to facilitate the transfer of genetic material, particularly when it is desirable to maintain the specific-pathogen free status of a herd.

The invention discloses a method for establishing an immunodeficiency mouse model. The method comprises the following steps of: (1) constructing DNA (deoxyribonucleic acid) which enables functions of an immune related gene in a mouse cell to be lost; (2) purifying mRNA (messenger ribonucleic acid), and cryopreserving in a super-purified water of RNAase free; (4) feeding a pregnant female mouse and a pseudopregnant female mouse under an SPF (specific pathogen free) level feeding condition; (5) taking a fallopian tube from the abdominal cavity of the pregnant female mouse in the step (4), and collecting fertilized ovum; (6) injecting the mRNA obtained in the step (3) into the pronucleus of the fertilized ovum in the step (5) by using a microinjection method; (7) transplanting the fertilized ovum obtained in the step (6) into the body of the pseudopregnant female mouse obtained in the step (4), and enabling the pseudopregnant female mouse to culture the second generation; (8) establishing a stable deficient mouse strain. The method has the advantages that a mouse model which completely does not have immunological rejection to allotransplantation can be obtained for testing.

The invention relates to a method for constructing a specific pathogen-free penaeid shrimp culture system, which comprises the steps of adopting comprehensive treatment of culture water for avoiding the entry of pathogens; introducing selected oocystis borgei into a penaeid shrimp culture pond for constructing a good microalgae community structure, and using a microalgae-controlled water environment to reduce the concentration of injurious factors, such as ammonia nitrogen, nitrite nitrogen and the like in a water body, improving the water quality, eliminating stress factors, improving the content of dissolved oxygen in the water and leading the environment of the water body to be in the good dynamic balance state for a long time. A system of the oocystis borgei, rhodopseudanonas palustris and phycomycete is utilized for inhibiting the growth of vibrios during the early stage, the middle stage and the late stage of culture; and schmackeria dubin is utilized to intake microalgae, thereby controlling the number of the microalgae in the water body and eliminating the adverse impacts of rapid proliferation of the microalgae on a culture environment. Penaeid shrimps have the advantages of specific pathogen-free property, rapid growth, strong disease resistance, strong stress resistance, short culture period, large and uniform formed size, high output, good investment efficiency and the like.

A production technology for SPF (specific pathogen free) chickens relates to the field of aviculture, solves the problems of low hatchability and egg laying performance, and poor environmental adaptability of the conventional SPF chicken production method, and comprises the following steps: sterilizing hatching eggs with formalin and fumigating the hatching eggs with potassium permanganate in an SPF chicken barrier system for 48 hours before hatching; 18 days after hatching, carrying out egg tray placing till chicks are hatched at the 21st day; feeding the chicks with a high-pressure sterilized SPF chicken brood fodder within 48 hours after the chicks are hatched; transferring the chicks into an adult chicken feeding room after the chicks are 42 days' old; conducting group transferring after the chicks are 17 weeks' old, wherein the chicks become adult chickens when 18 weeks' old, and an SPF adult chicken fodder, which is a full rate daily ration, is prepared according to the nutrition requirement of the chickens. Temperature and humidity are adjusted to be most proper and the size of an air door is enlarged properly according to hatching parameters, so that the hatching rate can be up to 98 percent; the fertilization rate of chicken groups mixed when the chickens are 17 weeks' old can be higher than 96 percent; a proper lighting program is adopted to ensure that the egg laying performance of chicken groups can be improved to be 88 percent maintaining for 2 weeks, and higher than 80 percent maintaining for 12 weeks.

The invention discloses a novel DHAV (duck hepatitis A virus) JS strain and application of the DHAV JS strain in duck virus hepatitis prevention and cure. The DHAV JS strain is obtained through the steps of separation culture, duck embryo neutralization test, PCR (polymerase chain reaction) detection of viruses and sequencing detection experiment of the viruses, and the strain is determined to be the DHAV serotype 3; and the microbial preservation number is CGMCCNo.6852. According to the invention, through taking the novel DHAV JS strain as the virus strain for producing vaccine, the novel DHAV inactivated vaccine is prepared through the steps of inoculating 10-day-old SPF (specific pathogen free) duck embryos, selecting the embryos died after incubating for 48 hours, collecting the embryo liquid, inactivating and emulsifying. The clinical application proves that the prepared inactivated vaccine is favorable in safety and conducive to prevention and control of the novel DHAV disease, can prevent the infection and outbreak of the novel DHAV in a targeted way, and can be in mass production and clinical application.

A method for obtaining an immunogenic strain useful for producing a vaccine against Coccidiosis comprises the cycle of infecting at least one first group of specific pathogen-free donor birds with oocysts from an Eimeria species. Blood is then collected from these donor birds, and is then used to infect a second group of specific pathogen-free birds. Oocysts are collected from the second group of birds. These oocysts are then multiplicated to complete the cycle. The cycle is then repeated using the multiplicated oocysts. After a total of about three cycles, a final antigen may be harvested and utilized as a source to generate oocysts for a vaccine.

The invention discloses an attenuated Salmonella pullorum and application thereof. By using gene knockout technology, the in vivo colonization gene and the main virulence gene of Salmonella pullorum are deleted, thus obtaining an dual-gene-deleted attenuated Salmonella pullorum SM091-DED strain; and the microbiological preservation number of the strain is CGMCC NO.4604. The virulence of the attenuated Salmonella pullorum disclosed by the invention is obviously reduced, and the in vivo colonization time is short after the attenuated Salmonella pullorum is inoculated into a host; the herd infection test shows that the attenuated Salmonella pullorum has no horizontal transmission capability; the attenuated Salmonella pullorum provides full cross protection for homotype and allotype high-virulence Salmonella pullorum; and after SPF (Specific Pathogen Free) chicks are immunized, the infection of the high-virulence strain can be effectively eliminated, and the herd infection can not be caused. Thus, the invention provides a safe, fine-immunogenicity and low-virulence Salmonellosis avium attenuated live vaccine for preventing Salmonellosis avium.

The invention relates to a feed for SPF (Specific Pathogen Free) breeding female rabbits in a pregnant period. The feed comprises the following components in parts by weight: 190-210 parts of corn flour, 90-110 parts of corn gluten feed, 355-375 parts of alfalfa meal, 190-210 parts of wheat bran, 1-2 parts of methionine the content of which is 98%, 2-4 parts of lysine, 4-6 parts of mountain flour for feed, 13-15 parts of calcium hydrophosphate, 2.5-3.5 parts of table salt, and 10 parts of premix. The invention also discloses a preparation method of the feed. The method comprises the following operation steps of: respectively weighting the raw materials; blending the weighted raw materials; uniformly stirring and packaging the raw materials; and sterilizing the packaged raw materials. According to the invention, the feed is scientific and reasonable in formula; the SPF breeding female rabbits in the pregnant period fed by the feed are high in farrowing rate, and the cubs are high in birth litter weight; and by adopting Co60 radiation sterilization, the content loss of nutrient substances is small, and the feed is easy to store.

The invention discloses a lactobacillus salivarius strain and application thereof. The lactobacillus salivarius strain (with number of HVRIC5) is obtained by carrying out bacteria reproduction and enrichment on wild chicken manure in an MRS liquid selective medium and carrying out isolated culture and inoculation and purification. The microbial collection number of the strain is CGMCC No.6787. Lactobacillus salivarius has stronger acid and bile salt resistance. Antibacterial experiments in vitro indicate that the strain has stronger antibacterial effects on Gram-negative escherichia coli, salmonella and Gram-positive staphylococcus, shows a broad-spectrum antibacterial effect, has the effect on promoting growth of SPF (specific pathogen free) chickens, can effectively resist infection with salmonella and avian infectious laryngotracheitis viruses when being fed to the SPF chickens and has the effect of effectively protecting the SPF chickens.

The invention discloses a feed used in the brooding period of SPF (specific pathogen free) breeding hens and a preparation method thereof. The feed is characterized by comprising the following raw materials in parts by weight: 590-610 parts of corn meal, 280-320 parts of bean pulp, 25-35 parts of wheat bran, 25-35 parts of fish meal, 0.5-1.5 parts of 98% of lysine, 1-2 parts of 98% of methionine,5-15 parts of limestone meal, 10-20 parts of calcium hydrophosphate, 2-3 parts of salt and 10 parts of premix. Compared with the traditional sterilization method, the preparation method is characterized by adopting a Co60 ray source for radiation sterilization and strictly limiting the radiation dose rate and radiation time. The feed subjected to radiation sterilization of the Co60 ray source haslow nutrient losses, good sterilization effect and low cost and is suitable for industrial production.

The invention discloses a method for quickly constructing an IBV (Avian Infectious Bronchitis Virus) reverse genetic strain, and belongs to the technical field of coronavirus reverse genetics. The constructing method comprises the following steps: quickly completing construction containing IBV genomic full-strength cDNA (Complementary Deoxyribose Nucleic Acid) clone by taking a BAC (Bacterial Artificial Chromosome) vector as a framework and applying an in-vitro homologous recombination technology, directly transfecting cells by a constructed recombinant plasmid, and transcribing in the cells, thus obtaining a transcript having infectivity; completing virus packaging; inoculating SPF (Specific Pathogen Free) chick embryo to a mixed solution of the cells and a culture medium and passing from generation to generation, thus obtaining the IBV reverse genetic strain. The constructing method disclosed by the invention has the advantages of simple operation and high positive cloning efficiency, the obtained IBV reverse genetic strain has passage stability, and an effective tool is provided for researching pathogenesis of the virus in vitro, developing a novel vaccine and the like; according to the method disclosed by the invention, transcription is carried out in the cells by utilizing a CMV (Cytomegalovirus) promoter added on a 5' terminal, and the rescue efficiency of the virus is greatly increased by utilizing an HDVR (Hepatitis Delta Virus Ribozyme) sequence added on a 3' terminal.

The invention relates to a detection kit and method for prawn infective virus, belonging to the technical field of marine organism pathogeny detection. The invention aims to provide a detection kit and method. The detection method is more sensitive and specific, and can shorten the reaction time and reduce the experimental steps. The detection kit comprises (1) an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) buffer tube, (2) a primer tube, (3) an enzyme tube, (4) a deionized water tube, (5) a negative control tube and (6) a positive control tube, wherein the RT-PCR buffer tube is filled with a 2*RT-PCR buffer; the primer tube is filled with infective muscle necrosis virus forward/reverse primers; the enzyme tube is filled with reverse transcriptase and DNA polymerase; the deionized water tube is filled with sterile deionized water; the negative control tube is filled with SPF (Specific Pathogen Free) prawn nucleic acid; and the positive control tube is filled with infective muscle necrosis virus cDNA (complementary deoxyribonucleic acid) positive clone. The detection method comprises (1) an RT-PCR reaction system and (2) an RT-PCR reaction procedure.

The invention provides universal CD8+T cell epitope polypeptide of a chicken IBV (Infectious Bronchitis Virus) S1 protein, and belongs to the field of gene and protein engineering. The epitope polypeptides are prepared by the following steps: screening 21 epitope polypeptides in accordance with binding motif sequences in amino acid sequences of the S1 genes of the IBV virus according to the binding motif sequences of haplotype chicken major histocompatibility complex (MHC) I-type molecules; then, taking lymphocytes of three constructed SPF (Specific Pathogen Free) chicken immunized by DNA (Deoxyribonucleic Acid) recombinant plasmids of the S1 genes containing different subtype IBVs, determining the capacity of the 21 polypeptides which induce chicken splenic lymphocytes to secrete interferon-gamma by using an ELIspot (Enzyme-Linked Immunospot Assay) method, and finally, screening to obtain four universal functional T cell epitope polypeptides of IBV, wherein the sequences are respectively shown in SEQ ID NO. 1-4. The four epitope polypeptides provided by the invention are combined in use to prepare a universal vaccine for IBV. The invention further provides a method for screening the epitope of the functional T cell of the S1 protein of the IBV.

The invention relates to a positive-serum and negative-serum standard substance of avian influenza virus H5N1 subtype Re-5 strain and a preparation method thereof. The preparation method comprises the following steps of: preparing a standard substance positive serum (strongly positive serum and weakly positive serum): intramuscularly injecting an avian influenza virus H5N1 subtype Re-5 strain oil-emulsion inactivated vaccine in a young or adult SPF (Specific Pathogen Free) chicken, and then collecting the serum of the SPF chicken and carrying out semi-finished product inspection, adding a proper stabilizing agent and then freeze-drying; preparing negative serum: taking the blood of the SPF chicken, separating serum, carrying out semi-finished product inspection, and freeze-drying; and carrying out a series of technical processes, such as finished-product inspection, uniformity inspection, stability inspection, calibration and value determination of the standard substance and the like to obtain the positive-serum and negative-serum standard substance. The standard substance is the fundamental guarantee for the accurate diagnosis of avian influenza virus H5N1 subtype, the immune monitoring of a H5N1 subtype Re-5 strain and the accurate evaluation on the vaccine immune effect of the H5N1 subtype Re-5 strain, thereby improving the prevention and control level of avian influenza. The standard substance is the basic guarantee for the diagnosis of avian influenza virus H5N1 subtype and the evaluation and the quality control of the inspection working of related products.

The invention discloses a lactobacillus salivarius strain and application thereof. The lactobacillus salivarius strain (with number of HVRIC4) is obtained by carrying out bacteria reproduction and enrichment on wild chicken manure in an MRS liquid selective medium and carrying out isolated culture and inoculation and purification. The microbial collection number of the strain is CGMCC No.6786. Lactobacillus salivarius has stronger acid and bile salt resistance. Antibacterial experiments in vitro indicate that the strain has stronger antibacterial effects on Gram-negative escherichia coli, salmonella and Gram-positive staphylococcus, shows a broad-spectrum antibacterial effect, has the effect of promoting growth of SPF (specific pathogen free) chickens, can effectively resist infection with salmonella and avian infectious laryngotracheitis viruses when being fed to the SPF chickens and has the effect on effectively protecting the SPF chickens.

The invention discloses a raising environmental system for specific-pathogen-free pigs. The raising environmental system is characterized in that a specific-pathogen-free (SPF) pig cage 5 is arranged in a clean raising room 16; the clean raising room 16 is provided with a raising room entrance 17 and windows 24, and a pedestrian path 15 is arranged in the clean raising room 16; air supply outlet 1 of a pig living quarter and an air supply outlet 3 of a pedestrian quarter are fixedly formed in a ceiling 2, and an exhaust outlet 6 of the pedestrian quarter and an exhaust outlet 11 of the pig living quarter are fixedly formed in a lateral lower side of a wall; an upper space of the pig living quarter can be separated from the pedestrian quarter by an isolation curtain 4; an isolation door 13 driven by a lifting device 14 is arranged in the pig cage 5; the pig cage 5 is provided with an excrement leaking floor 12, a sewage guide plate 8 and a disinfectant fluid sprayer 9 are arranged below the excrement leaking floor 12, and the sewage guide plate 8 is connected with a negative-pressure suction device 7 by a blowdown pipe 10; a disinfectant fluid generator 30 is connected with the disinfectant fluid sprayer 9 by a disinfectant fluid pipe 31. The raising environmental system has the advantages that cross infection of various pathogens can be prevented, the pigs can be raised in a separated or group manner as needed, and excrement can be timely thoroughly cleaned and disinfected.

The invention relates to a cultivation house for test animal of genus Macaca without specific pathogen. Interior region of the cultivation house is composed of a working room, an observation room, a storehouse, a corridor, and several cultivation rooms; periphery of the cultivation room is sequentially installed with iron wire protective net, nanometer protective net and anti-rust angle iron keel from inside to outside, to completely enclose the cultivation room from exterior open part of the cultivation house; and device for supplementing preventative and health promoting/conditioning liquid is installed above the working room. Through the cultivation house structure, propagation of pathogens (bacteria, parasite and virus) carried by rodents, insects, birds and other animals can be effectively controlled; infection rate of cultivated SPF class test machin and rhesus due to exterior reasons (bacteria, parasite and virus) is reduced by 99%; and by adding the device for supplementing preventative and health promoting/conditioning liquid, uniform supply of microelements, vitamins and minerals can be satisfied to form favorable guarantee for test monkey health.

The present invention relates to a method of producing a SPF (specific pathogen free: SPF) hamster for the preparation of a variety of biological products. More particularly, the present invention relates to a method of producing the SPF hamster for the preparation of a variety of biological products, that is free of microbiological contaminants such as parasites, bacteria, and exogenous- and endogenous viruses, etc. with the potential to cause diseases. Since the SPF hamster produced according to the method of the present invention carries no contaminants such as parasites, bacteria, and exogenous- and endogenous viruses, etc., the primary cells originated from said SPF hamster and the biological products using the same do not induce any opportunistic infection by microbial contaminants that can be present in animal cells used for existing biological products such as vaccine, etc. Therefore, the SPF hamster produced according to the present invention can be safely used for preparing a variety of biological products that are applied to a human.

The invention relates to preparation and application of a newcastle disease-infectious bursal disease VP2 subunit bivalent vaccine. The preparation comprises the following steps of performing egg inoculation on a newcastle disease Clone30 strain to obtain blastochyle, and inactivating; mixing with chicken infectious bursal disease VP2 protein liquid of bacillus coli in a certain proportion; adding mineral oil adjuvant, and emulsifying to obtain bivalent vaccine; immunizing a 21-day SPF (specific pathogen free)chick with the bivalent vaccine; performing toxicity attack on newcastle disease virulent strain isolated in Beijing and infectious bursal disease virulent BC6-85 respectively after 21 days of immunization, results indicate that light bursa fabricius symptom appears on a chick in 20 chicken in the immunized group, and the rest chicken does not have abnormality; and 10/10 chicken are attacked and dead in the newcastle disease control group, and 8/10 chicken in the infectious bursal disease virus attack control group are attacked.

The invention provides an anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody which can be specifically bound with schistosoma japonicum Sjp40. The anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody is capable of identifying the schistosoma japonicum Sjp40 through antigen specific binding activity detection thereof; the OD value of the anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody is decreased as the antibody dilution is reduced; the anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody has dependence on antigen concentration; the antibody is obtained by a method comprising the following steps of: (1) collecting eggs of healthy SPF (Specific pathogen Free) grade laying hen which is primarily immunized with purified Sjp40 antigen previously, and primarily extracting IgY from the eggs by the method of diluting ammonium sulfate precipitate with water; (2) further purifying the IgY orderly through a dialysis bag and a DEAE-FF (Diethylaminoethyl-Fast Flow) anion exchange column; (3) performing analysis and identification on the IgY through polyacrylamide gel electrophoresis, Western blotting and ELISA (Enzyme Linked Immunosorbent Assay), wherein the antibody is a chicken egg-yolk immune globulin (IgY) special for the schistosoma japonicum Sjp40; and therefore, a new idea is provided for early diagnosis and prevention and treatment of the schistosoma japonicum.

Transgenic non-human mammals comprising a DNA with an exogenous IL-18 gene incorporated in it such that the exogenous IL-18 gene is skin-specifically expressed, or their offsprings.

The transgenic non-human mammals according to the present invention spontaneously develop atopic dermatitis under specific pathogen-free conditions, and therefore, are useful as disease model animals. Use of the transgenic non-human mammals according to the present invention makes it possible to develop drugs for preventing or treating atopic dermatitis by natural immunity and also to elucidate the onset mechanisms of atopic diseases.

The invention relates to a special transportation box for specific pathogen free (SPF) stage mice and belongs to the technical field of life science. The special transportation box for SPF stage mice comprises a box body, a box cover and a stainless steel metal screen, wherein the box cover is covered on the box body, the stainless steel metal screen is arranged in the box body, a plurality of ventilation windows are respectively arranged at two sides of the box body and on the box cover, and the ventilation windows are formed through melting filter films and polypropylene plastics and are integrally formed with the box body and the box cover. Through adopting a scheme that a plurality of ventilation windows which are formed through melting filter films and polypropylene plastics and integrally formed with the box body and the box cover are respectively arranged at two sides of the box body and on the box cover, the sealing performance is good, and the environment in the box body can be effectively controlled from being polluted.

The invention provides a method for separating acian metapneumovirus, which adopts tracheas and lung tissues mixed cells of specific pathogen free (SPF) chick embryos which are incubated for 17-19 ages in days to cultivate the acian metapneumovirus. The method for separating the acian metapneumovirus successfully improves acian metapneumovirus (AMPV) vitro culture conditions through a separation and reproduction method of a same culture system and two kinds of mixed cells based on a traditional method that a clinical development sample is separated and cultured into a kind of sensitive cell at one time, obviously improves separation rate of AMPV, and reduces reproduction times.

The invention relates to the field of preparation of pharmaceuticals, in particular to application of Bacteroides thetaiotaomicron VPI-5482 in the preparation of pharmaceuticals for treating or preventing obesity. C57/BL6 male SPF (specific pathogen free) mice of the same litter are randomly divided into a control group and a bacterial filling group. The control group is subjected to aseptic PBS lavage; the bacterial filling group is subjected to Bacteroides thetaiotaomicron VPI-5482 suspension lavage test once a day; feeding with high fat food is performed for 7 weeks of testing, during this period, body weight increase rate of the mice is recorded, and after testing, fat content and the like are detected. Testing shows that the Bacteroides thetaiotaomicron VPI-5482 is very effective in controlling body weight and fat, inguinal subcutaneous fat content is decreased by 35.5%, and near-epididymal fat content is decreased by 23.5%.

The invention discloses a feed for specific pathogen-free (SPF) chicken in the peak laying period (daily age of 127-420) and a preparation method for the feed. The feed is characterized by being prepared from the following raw materials in part by weight: 620 to 640 parts of corn flour, 190 to 210 parts of bean pulp, 15 to 24 parts of wheat bran, 35 to 45 parts of fish meal, 1 to 2 parts of methionine with the purity of 98 percent, 70 to 90 parts of mountain flour, 10 to 14 parts of calcium hydrophosphate, 2.5 to 3.5 parts of table salt and 10 parts of premix. Different from the traditional sterilization method, the invention has the advantages that: a Co60 radiation source is adopted for sterilization, and a radiation dose rate and radiation time are strictly limited. The feed for the SPF chicken, which is sterilized by the Co60 radiation source, has small loss of nutritional components, is well sterilized and low in cost, and is suitable for industrial production.