65 results about "Chicken serum" patented technology

The invention aims at providing an I-colony fowl adenovirus 4 strain, which is preserved with preservation number of CCTCC No. V201541. The I-colony fowl adenovirus 4 YBAV-4 strain disclosed by the invention is excellent in specificity and immunogenicity; a specific precipitation line does not appear in specific positive serum chicken SPF chicken serum such as infected cell sap and egg drop syndrome resisting virus, chicken infectious bursal disease virus, Newcastle disease virus, chicken infectious laryngotracheitis virus, chicken Marek's disease virus, avian influenza and the like, while an obvious specific precipitation line appears in I-colony fowl adenovirus 4 specific serum. The strain disclosed by the invention, as a vaccine strain which is good in manufacturing effect, is capable of preventing chicken hydropericardium syndrome, and the strain is applicable to identification of virus serotype and investigation on epidemiology.

The invention discloses a preparation method and application of Newcastle disease virus infected immune complex vaccines. The preparation method comprises the following steps of: (1) preparing antigens and inactivated vaccines by using Newcastle disease virus Shanghai strains as seed viruses; (2) transferring antibodies in chicken blood serum to yolk to form yolk antibodies IgY; and (3) mixing the prepared inactivated vaccines and the yolk antibodies IgY in equal volume, and hatching to obtain the immune complex vaccines. By combining passive immune (vaccine) and active immune (specific antibody) measures, animals are protected from the infection of pathogenic microbes in time for a long time.

The invention discloses a method and a special culture medium for subculturing chicken embryonic stem cells for a long time. The method is characterized by comprising the following steps of: isolating cells of area pellucida of X-stage blastoderm, dispersing into single cells or small cell masses through mechanical blowing and beating, inoculating on an STO feeder layer, culturing the chicken embryonic stem cells in the special culture medium at the temperature of between 37 and 38 DEG C, and subculturing once every 3 to 5 days, wherein the special culture medium comprises 600 to 900mL of conditioned medium, 50 to 150mL of fetal calf serum, 5 to 20mL of chicken serum, 5 to 25mL of one or a mixture of more of non-essential amino acids, 0.146 to 0.292g of L-glutamine, 6 to 14mu L of beta-mercaptoethanol, 1 to 2*10<6> IU of mouse leukemia inhibitory factor, 10 to 50mu g of alkaline fibroblastic growth factor and 5 to 20mu g of stem cell growth factor; and fixing the volume of the ingredients to 1,000mL by using a dulbecco's modified eagle medium (DMEM) (high glucose), and regulating the pH value to 7.2 to 7.5. The conditioned medium is obtained by the steps of: culturing BRL-3A cells until the cells are converged, collecting supernatant of the cultured cells, performing centrifugal separation, and filtering by using a filter membrane to obtain the conditioned medium. Compared with the prior art, the method has the advantages that: the long-term subculture of the chicken embryonic stem cells can be realized, the proliferation of the cells is quick, and the cloning yield is high.

The invention discloses a traditional Chinese medicine feed additive and an application thereof. The traditional Chinese medicine feed additive disclosed by the invention is an animal feed additive, and the active ingredients of the animal feed additive comprise the following traditional Chinese medicine raw materials in parts by weight: 10-30 parts of lycium barbarum, 10-30 parts of wolfiporia cocos, 30-60 parts of fructus Hippophae, 30-60 parts of flos Lonicerae and 30-60 parts of polygonatum sibiricum red. The animal feed additive disclosed by the invention has the following advantages of reducing the content of cholesterol in eggs, increasing the contents of selenium, ferrum and zinc, increasing the contents of total protein in chicken serum, the content of globulin and the convent of alkaline phosphatase and increasing the content of complement C-4 in the chicken serum.

The invention relates to a method of utilizing yolk antibody instead of serum antibody to evaluate infection state of breeder flock. The method of the invention comprises the following steps: collecting chicken serum and eggs, respectively; respectively detecting antibody levels of serum and yolk by REV, REOV, ALV-J, ALV-AB, CAV and IBDV enzyme linked immunosorbent assay (ELISA) kits from IDEXX company; comparatively analyzing serum and yolk antibody OD value correlation coefficient by SPSS Statistics 17.0 to obtain correlation coefficient of an optimum dilution factor of yolk antibody and a serum antibody OD value according to ELISA detection. The detecting result further indicates that the positive and negative result consistent rate of yolk sample and the serum sample at the optimum yolk dilution factor is 96%-100%. The result of the invention indicates that the serum samples can be replaced by yolk when various infection states of breeder flock are detected by ELISA kits provided by IDEXX for detection with serum samples, so that the infection state of breeder flock can be detected by detecting yolk antibody level.

The invention discloses a visual protein chip for detecting serum antibody of new-castle disease virus of chickens, infectious bronchitis virus of chickens, avian influenza virus and infectious bursal disease virus of chickens , which is prepared by the following steps: purifying and diluting whole proteins of the four virus respectively; pointing samples of the positive control serum, the negative control serum and the four virus proteins onto a chip carrier respectively; drying, fixing, sealing and washing the samples to obtain the visual protein chip. The visual protein chip uses the purified whole proteins as capturing antigens to detect the virus-specific antibodies in chicken serum so as to simplify the preparation technology and reduce the production cost, and the visual protein chip has better specificity but no cross, has high reliability of results and has the advantages of quickness, simplicity and convenience, high sensitivity, good specificity and the like. When the serum is diluted by 6,400 times, the visual protein chip still can detect the antibodies, the sensitivity is 400 times of that of the prior AGP detection method. According to the detection to serum samples in-place, the detection rate of the visual protein chip is higher than the proir AGP method remarkably.

The invention discloses an ELISA (enzyme-linked immunosorbent assay) detection method for identifying fowl adenovirus group I (FAVI) infection. The method is characterized by taking an FAVI 100K recombinant protein as the envelop antigen, chicken serum as the detection sample and horse radish peroxidase labeled goat anti-chicken IgG as the enzyme-labeled antibody to detect the antibody generated through FAVI infection. The method is strong in specificity, short in time and low in cost, is simple to operate, can be used for mass detection, can effectively get rid of interference of immunity of FAVI inactivated vaccines and can specifically detect FAVI infection, thus distinguishing FAVI infected animals from FAVI inactivated vaccine inoculated animals and providing a diagnostic tool with actual value for eliminating FAVI in China.

The external cultivating method of primordial bird germ cell relates to transgene, animal cloning, rare animal preservation, tissue and cell engineering and other science fields. After it is inoculated to the CEF raising layer, the primordial bird germ cell is cultivated in liquid high-sugar DMEM cultivation medium with added calf serum in 10 úÑ, chicken serum in 2 úÑ, L-glutamine in 2mmol/L, sodium pyrovate in 1 mmol/L, beta ¿Cmercaptoethanol in 5.5í‡10-5 mol/L, optional amino acid 10 microliter/ml, hSCF in 15-20 ng/ml, mLIF in 10 U/ml, bFGF in 10-15 ng/ml, hIL-11 in 0.04 ng/ml, IGF in 10 ng/ml, and gentamycin sulfate in 100 U/ml. The present invention has simple operation and high repeatability, and lays the foundation of the relevant research.

The invention discloses a nanobody capable of resisting H9N2 subtype avian influenza viruses, a preparation method and an application. The amino acid sequence of the nanobody is as shown in SEQID NO:1. The nucleocapsid protein of the H9N2 subtype avian influenza viruses is expressed through a prokaryotic expression technique, then the nucleocapsid protein is used as an immunogen for immunizing Bactrian camels, a phage library is constructed, then a nanobody capable of resisting H9N2 nucleocapsid protein is obtained through screening by a phage display technique, then the nanobody and horse radish peroxidase (HRP) are subjected to fusion expression, and a fusion protein of the nanobody and the HRP is successfully prepared. When the fusion protein is applied to detection of antibodies capable of resisting H9N2 subtype avian influenza viruses (AIV) in chicken serum, the inventor finds that a detection method constructed through the fusion protein is high in sensitivity, and has the advantages that the operation is simple, secondary antibodies do not need to be used, and time consumed for detecting samples is short.

The invention belongs to the technical field of cell isolation and cultivation. In order to solve the problems that the chicken small intestinal epithelial cells are difficult to isolate and culture in vitro at present, the existing isolation process is troublesome and time-consuming, the obtained cells are low in purity and the like, the invention provides isolation and primary culture methods of the chicken small intestinal epithelial cells. The methods comprise the steps of taking chick embryo small intestines, cleaning the small intestines to remove mesenteries, digesting small intestine tissues by protease to obtain the chicken small intestinal epithelial cells, performing adherent difference culture on the cells, removing adherent parenchyma cells, collecting intestinal epithelial cells, inoculating by taking the inoculum density being about 6.5X105/mL for culturing, the chicken small intestinal epithelial cells obtained by culturing is maximum in quantity, low in death rate, and best in growing status. The primary culture is performed by adopting a culture solution in which chicken serum is added, and the growing status of the obtained cells is better. The chicken small intestinal epithelial cells which are good in growing status, normal in shape, high in activity are successfully cultured, the primary and isolation culture methods of the chicken small intestinal epithelial cells are successfully established, and the technical support is provided for follow-up tests.

The present invention relates to an inert carrier Escherichia coli and its potential application, the Escherichia coli isolated strain has the inert carrier property, and is expected to be developed into a carrier strain for an indirect agglutination test. The inert carrier Escherichia coli is preserved in the China General Microorganisms Collection and Management Center (CGMCC), and a preservation address is Beijing, China, a preservation number is CGMCC No. 17339, a preservation date is March 18, 2019, a classification name is Escherichia coli, and a strain code is SE1. The Escherichia coli isolated strain is from a healthy flock, the bacteria number at a higher working concentration does not cause any macroscopic agglutination reaction with various chicken serums of different genetic backgrounds, and does not generate non-specific agglutination reaction with chicken source serum, so that the Escherichia coli is called as an inert carrier Escherichia coli. The application provides an inert carrier strain and its potential application for developing a simple and rapid indirect agglutination assay.

The invention belongs to the technical field of agricultural breeding, and particularly relates to a nutritional broiler chicken breeding method which includes the stages of chicken house management before chick entering, 1-7-day-old management, 8-14-day-old management, 15-26-day-old management, 27-34-day-old management and 35-day-old to slaughter management. Compared with the prior art, the breeding method has the advantages that the breeding method is further improved based on existing breeding, a chick growth system can be rapidly perfected, the absorption and utilization rate of broiler chicken for nutritional substances in feed is increased, the concentration of the nutritional substances in broiler chicken serum is increased, the meat of the bred broiler chicken is delicious and high in nutrition, nutritional agents of the feed do not remain in bodies of the broiler chicken, toxic and side effects are avoided, chicken nutrition can be increased, high slaughtering rate of the broiler chicken is ensured, and the breeding method is suitable for popularization and use.

The invention belongs to the field of livestock raising and discloses a method for bringing forward the sexual maturity of a cock. The method for bringing forward the sexual maturity of the cock comprises the following steps that before the hatching, the small end of a hatching egg is sterilized, a hole is slightly drilled in the sterilized part by a toad spinal cord puncture needle, a trace sample feeder is used for introducing 100 muL 0.5 to 2ng/muL kisspeptin-10 injection reagents into egg white through the drilled hole, and an opening is sealed with paraffin wax. The kisspeptin-10 is used for processing the hatching chicken egg for the first time, in addition, the processing concentration of kisspeptin-10 is optimized, results show that when the method provided by the invention is used for Kp-10 (kisspeptin-10) egg injection, the body weight of the cock can be obviously increased, the sperm density of the cock is improved, the sexual behavior of the cock is increased, and the obvious influence is realized on the T content in the blood serum of the 14-week and 24-week cock.

The invention discloses a haemophilus paragallinarum B strain fermentation medium, a preparation method thereof and application thereof, and belongs to the field of agricultural microorganism. The haemophilus paragallinarum B strain fermentation medium is prepared from 5 to 100 g/L of casein peptone, 0.5 to 80 g/L of yeast powder, 0.5 to 80 g/L of polyvalent peptone, 0.5 to 20 g/L of sodium chloride, 0.5 to 20 g/L of sodium glutamate, 0.5 to 30 g/L of glucose, 0.5 to 30 g/L of fish meal peptone, 5 to 100 mL/L of chicken serum, 0.5 to 50 mL/L of microelement solution and 1 to 30 mL/L of NAD (Nicotinamide Adenine Dinucleotide) solution. The haemophilus paragallinarum B strain fermentation medium disclosed by the invention contains all nutritional ingredients which are needed for growth of haemophilus paragallinarum B strains, growth and metabolism of the strains are facilitated, the adaption period of thallus is shortened, the adaptive capacity of the thallus after inoculation is strong, the fermentation period is short, the viable bacteria quantity during harvesting is high, and the haemophilus paragallinarum B strain fermentation medium is suitable for large-scale application and has a wide prospect.

The invention discloses a method for culturing chicken embryonic germ cells and a special culture solution thereof. The culture solution is a high sugar DMEM solution containing the following solutes: 3.5 to 4.5 g/L of glucose, 1.5 to 2.5 mM of L-glutamine, (4.5 to 5.5) x 10<5> M of beta-mercaptoethanol, 1 to 1.5 mM of hydroxyethyl piperazine ethyl sulfuric acid, 100 to 120 IU/ml of leukemia inhibitory factor, 4 to 6 ng/ml of stem cell growth factor, 10 to 12 ng/ml of basic fibroblast growth factor, 10 to 12 ng/ml of insulin-like growth factor-1, 10 to 15 percent of fetal bovine serum, 1 to 3 percent of chicken serum, 1 to 1.2 percent of chick embryo extract, and antibiotic. The method for culturing the chicken embryonic germ cells is to culture the chicken embryonic germ cells on a chicken embryonic fibroblast culture layer. Through the reformation and the modification of EG cells, new varieties of chicken can be effectively cultured.

The invention relates to a feed with medicinal functions, and concretely relates to a Chinese herbal medicinal premixed feed for preventing avian influenza of chickens, and a preparation method thereof. The Chinese herbal medicinal premixed feed comprises, by weight, 2-4 parts of honeysuckle flower, 2-3 parts of Herba Schizonepetae, 2-3 parts of Caulis Perillae, 2-3 parts of Radixet Rhizonma Ephedrae, 2-3 parts of Divaricate Saposhnikovia Root, 2-3 parts of white peony root, 2-4 parts of Radix Platycodonis, 2-4 parts of dry ginger, 2-3 parts of Cyrtomium fortunei, 6-7 parts of isatis root, 6-7 parts of Folium Isatidis and 60-65 parts of rice bran. A result of chicken serum examination of chicken flocks fed with the Chinese herbal medicinal premixed feed in a Hunan Polytechnic of Environment and Biology lab shows that the H1 antibody average level is higher than 410 g<2>.

The invention discloses a chicken skin yellowness breeding method. The method comprises the following steps: 1) determining cloaca skin color yellowness, chicken serum xanthophyll concentration and chicken serum beta-carotene concentration of a chicken to be detected; 2) solving a yellow skin breeding value according to a formula S = 0.5 * [B + (5 * C)] + 0.5 * (50 * A); and 3) retaining the to-be-detected chicken with the S value greater than 600. According to the chicken skin yellowness value breeding method, the two methods of detecting the skin yellowness value through the colorimeter and detecting the serum yellow pigment concentration value through the liquid chromatograph are adopted to cooperatively select the chickens with the high skin yellowness value and the low skin yellowness value, then large-scale breeding is conducted, the influence of the external environment on the skin yellowness value is reduced, the skin color is yellow and uniform, and market requirements are met.

The invention belongs to the technical field of biological engineering, and relates to a recombinant chicken osteocalcin maturation protein monoclonal antibody and application thereof. The recombinant chicken osteocalcin maturation protein monoclonal antibody is secreted by hybridoma cell 3H4 with the preservation number of CGMCC No.4502. The preservation number of the hybridoma cell 3H4 which ispreserved in the china general microbiological culture collection center at December 23rd, 2010 is China general microbiological culture collection (CGMCC) No. 4502. The hybridoma cell system 3H4 canstably secrete the recombinant chicken osteocalcin maturation protein monoclonal antibody, wherein the antibody is good in specificity and can be combined with the specificity of the chicken osteocalcin protein to assay the chicken osteocalcin protein. The chicken osteocalcin assay reagent prepared by the antibody and particularly the chicken osteocalcin enzyme-linked immuno sorbent assay (ELISA)kit can be used for assaying the content of the chicken serum osteocalcin, thereby being good for monitoring the bone growth condition, preparing the reagent for diagnosing the chicken bone disease, and diagnosing the other diseases which can cause the abnormal change of the BGP.

The invention discloses H5 subtype avian influenza virus HA protein B cell epitope polypeptide and application thereof. An H5 subtype AIV (avian influenza virus) HA amino acid sequence is used as the template, the epitope polypeptide is synthesized on a solid-phase carrier in situ, each polypeptide contains 15 amino acids, and 14 amino acids of each previous polypeptide and the corresponding next polypeptide overlap. The synthesized H5 AIV HA epitope is allowed to have reaction with known H5 subtype positive serum, and one epitope polypeptide (H5-1) which can have specific reaction with H5 AIV chicken serum. The epitope polypeptide is used for detecting H5 subtype avian influenza viruses, and the detection result shows that the ELISA method built by the epitope polypeptide is suitable for detecting a 1:200 diluted serum sample. A coincidence rate test proves that the detection result of epitope polypeptide on H5 AIV negative serum is negative, the detection coincidence rate of samples with the hemagglutination inhibition values ranging from log2 to log4 is 87.6%, and the detection coincidence rate of samples with the hemagglutination inhibition values being log5 or above log5 is 98.3%.

The invention relates to a separation and purification method and an identification method of chicken apolipoprotein AI, and relates to the technical field of biology. The separation and purification method comprises the following steps: collecting chicken serums; preparing serum high-density lipoprotein; defatting HDL (High Density Lipoprotein); dissolving the defatted HDL; dialyzing and concentrating a defatted HDL solution; dialyzing the HDL solution by using a urea-10mmol/LTris-HCl-1mmol/LEDTA mixed solution of 6 mol/L to remove an organic solvent contained in the HDL solution, and then concentrating polyethylene glycol 6000 of 500 g/L to 2-3 mL to enhance the concentration of the HDL solution. The separation and purification method disclosed by the invention is simple and lays the foundation for the measurement and further research of the apo AI.

The invention provides an ELISA method for detecting the titer of a cLHRH antibody in chicken serum. The method comprises the following steps: preparation of a coating antigen, coating, washing, blocking, addition of a sample, addition of an enzyme-labeled antibody, color development, termination of a reaction, and titer determination. According to the invention, synthetic peptide containing dualepitopes and polylysine is designed as the coating antigen, and the ELISA method containing the coating antigen is established for the first time for determination of the titer of the cLHRH antibody in chicken serum. The coating antigen is cLHRH-Lys10-cLHRH; and the titer determination is implemented by using an end-point dilution method or a standard curve method. The method provided by the invention can be used for detecting the titer of the cLHRH antibody in chicken serum, and is accurate, reliable and stable in detection results.

The invention discloses a preparation method of chicken interferon lyophilized preparation and belongs to the field of animal vaccines. The preparation method of the chicken interferon lyophilized preparation comprises six steps as follows: healthy chicken blood is collected sterilely and is subjected to centrifugation at the temperature of 10 DEG C for 20 min, stratum intermedium leukocytes are sucked, washed twice with NH4Cl with the mass percentage of 0.85% and then subjected to centrifugation, precipitated leukocytes are collected, NH4Cl is removed, an Eagle's MEM mixed culture solution containing 5% of chicken serum is added in a volume ratio of the leukocytes to the culture solution being 1:15 for suspension, the chicken leukocytes are diluted to be in a range from 10*10<7>/ml to 25*10<7>/ml, and a chicken leukocyte suspension is prepared. The preparation method of the chicken interferon lyophilized preparation has the characteristics that the long-acting chicken virus interferon preparation is prepared, chicken are injected with the preparation every 10 days, and the preparation has good effect on chicken virus diseases.

The invention discloses separation purification and identification methods of a chicken apolipoprotein A II, and relates to the technical field of biology. The separation purification method comprises chicken serum collection, crude HDL preparation, HDL crude product degreasing, degreased HDL dissolving, dialysis and concentration of the degreased HDL solution, extraction of the apolipoprotein A II, chromatography with a DEAESepharoseCL-6B column, and chromatography with a SephadexG-150 column; according to the method, the SephadexG-150 chromatography column is prepared, then an apoA II-containing protein solution passing through the DEAESepharoseCL-6B is added on a gel column, when the protein solution is parallel and level to a colloid, elution is carried out by using an eluant, concentration is carried out with sucrose, then the product is preserved for standby analysis. The method is simple, is strong in practicability, helps people further understand physiological functions of the apoA II, and can be used in epidemiological and clinical studies.

The invention discloses an application of chicken Marek's disease virus envelope glycoprotein gB protein in preparation of an indirect ELISA kit for detecting a chicken Marek's disease virus antibody,and provides the indirect ELISA kit for detecting the chicken Marek's disease virus antibody established by using the MDV envelope glycoprotein gB protein. The kit makes up the detection defects by aiming at the chicken MDV antibody, and has the advantages of simpleness, stabilization and high efficiency, the kit is used for rapidly detecting the MDV virus antibody in a clinical chicken serum sample at an early stage, can be used for monitoring the chicken immune antibody, and is an effective method for large-batch detection, and has the advantages of high specificity, sensitivity, and reappearance. Compared with a current traditional method, the detection effect is better.