35 results about "Head Kidney" patented technology

The invention relates to an epinephelus coioides interferon IFNgamma1 gene, a protein coded by the gene and an application of the protein in preparing a novel immunopotentiator. According to the invention, an opened reading frame of the IFNgamma1 gene is cloned from head-kidney RNAs (Ribonucleic Acid) of epinephelus coioides through a method of screening a genome database and designing specific primer amplification. The nucleotide sequence of the gene is shown as SEQIDNO:2. The amino acid sequence of the protein coded is shown as SEQIDNO:1. Through initial verification, the epinephelus coioides interferon IFNgamma1 protein can induce associated expression of immunogenes and can be developed to natural immunopotentiators or immunologic adjuvants for fishes.

The invention discloses a grass carp IFN-γ2 gene, which has the nucleotide sequence shown in SEQ ID NO: 1 in the sequence table. The invention also discloses a recombinant protein expressing the grass carp IFN-γ2 gene, which has the amino acid sequence shown in SEQ ID NO: 2 in the sequence table. The recombinant protein can promote the activation of grass carp head kidney macrophages and up-regulate the expression of immune-related genes in vitro. Injecting the recombinant protein into grass carp can significantly reduce the mortality rate of grass carp infected with Flavobacterium columnar, improve the immune protection rate of grass carp, regulate the immune index of grass carp, and enhance the ability of grass carp to resist bacterial infection.

The present invention provides a method for the isolation and primary culture of grass carp dendritic cells, comprising the following steps: Step 1, the separation of grass carp spleen and head kidney: the grass carp is anesthetized to death, the spleen and head kidney are taken out, and the attached connective tissue is removed and adipose tissue and soaked; step 2, preparation of grass carp spleen and head kidney single cell suspension: continuous sieve filtration to obtain cell suspension; step 3, separation of grass carp spleen and head kidney mononuclear cells: Ficoll separation, after centrifugation Absorb and wash the cells of the buffy coat; step 4, static culture: adjust the concentration of monocytes, and place them in an incubator for culture; step 5, enrichment and purification of grass carp dendritic cells: collect suspension cells after culturing for a period of time, and use Grass carp dendritic cells were purified by density gradient centrifugation. The method can effectively prepare a large amount of grass carp dendritic cells, and the purity of the obtained grass carp dendritic cells can reach more than 80 percent; the operation process is simple and clear, and the time consumption is short.

A method for preparing chromosomes of red-spotted grouper adult fish belongs to the field of cell biology, and it includes pre-treatment of materials, preparation of kidney cell suspension, hypotonic treatment, fixation, dripping and staining; the pre-treatment of materials is Using the phytohemagglutinin injection method in vivo, inject PHA at the base of the pectoral fin of the experimental fish at a dose of 15 μg / g fish body weight, and inject colchicine solution at a dose of 2 μg / g fish body weight after 24 hours, and after 2.5-3 hours , take out the head kidney. The present invention adopts the local method of one-time injection of phytohemagglutinin, increases the injection concentration at the same time, can obtain good effect after 24 hours, and saves time. In the present invention, due to the characteristics of tissue cells of the red-spotted grouper, it is difficult to obtain complete cells when the hypotonic treatment is performed according to the prior art. Therefore, the present invention creatively changes the prior art and obtains a clear chromosome picture.

The invention discloses a grass carp bacterial septicemia and red skin disease combined vaccine and a preparation method thereof, using β-propiolactone as an inactivator to develop safe and efficient grass carp Aeromonas hydrophila septicemia and Pseudomonas aeruginosa Bacterial erythroderma dual white oil inactivated vaccine can effectively control the occurrence of bacterial septicemia and erythroderma in grass carp. After immunizing grass carp, it can stimulate the expression of IgM, IL-1β, IFN, Mx, C3 and TLR3 immune-related genes in the spleen and head kidney tissue of the fish, and the vaccine can produce a relative immune protection rate of 80-90% for grass carp , the relative protection rate can still maintain 50% 1 year after immunization; the invention is easy to operate, safe and effective, and two diseases can be prevented by one immunization, which not only reduces the use of antibiotics and chemical drugs, but also reduces the impact on the breeding environment and ecological environment, etc. The pollution and destruction of the external environment ensure the food safety and water ecological safety of grass carp.

The invention discloses a method for improving the streptococcus iniae infection resisting capability of GIFT tilapia in high-density culture. The method comprises the steps of preparing of compound Chinese herbal medicine, adding, feeding and breeding management. Compared with the prior art, the method has the following advantages: (1) the immunological protection of the GIFT tilapia in high-density culture can be effectively improved, and the survival rate of GIFT tilapia infected with streptococcus iniae is improved; (2) the compound Chinese herbal medicine added in a feed belongs to environment-friendly feed additives, and has less negative effect on fish and water; (3) the blood and head-kidney immunological stress response can be effectively enhanced, and the liver stress injury is reduced; (4) the adopted technological steps of the method are simple, the Chinese herbal medicine is low, and easy to obtain and preserve, and can be well absorbed and utilized by a fish body, and excrement of the GIFT tilapia does not impact the water quality.

The invention discloses a system for characterizing macrophage phagocytosis microorganisms of pompanos. The pompanos are dissected and separated through a surgical knife and tweezers to obtain heads and kidneys; the heads and the kidneys are abraded through a cell sieve to obtain cell suspension containing macrophages, and the macrophages of the heads and the kidneys are obtained through a Percoll solution density gradient centrifugal tube. The survival amount of cells cultured in vitro through a trypan blue dyeing manner is counted, and the survival condition in one week is detected. The system is used for carrying out a phagocytosis experiment, and phagocytosis conditions of nocardia, yeast and the like of the pompanos by the head and kidney macrophages of trachinotus ovatus are determined, and a phagocytosis function is observed. An experiment result of the system shows that the macrophages of the pompanos can be extracted very well, and the survival condition and the phagocytosis condition of the macrophages of the pompanos can be observed very well. The system disclosed by the invention lays a foundation for further researches on physiology, pathology and toxicology of the macrophages of the pompanos.

The invention provides a tilapia feed additive for reducing proinflammatory response and enhancing immune response, a preparation method and application thereof, and belongs to the technical field of aquaculture. Tilapia feed additives for reducing pro-inflammatory response and enhancing immune response, comprising the following components in parts by weight: 25-35 parts of Acanthopanax stems, 15-25 parts of loquat leaves, 15-25 parts of Tripterygium wilfordii, 10-35 parts of hawthorn 20 parts and 10-20 parts of mulberry leaves. The invention also provides a tilapia feed containing the tilapia feed additive. Tilapia feed additive or the application of said tilapia feed in tilapia culture, said application can effectively reduce the pro-inflammatory response of tilapia head kidney tissue and spleen tissue, enhance its immune response ability, thereby Improve its ability to resist streptococcal infection.

The invention discloses a pseudosciaena crocea head kidney cell line and a construction method thereof, relating to cell lines. The pseudosciaena crocea head kidney cell line is a pseudosciaena crocea head kidney cell line LYCK with the preservation number of CCTCC NO: C2013113. The construction method of the pseudosciaena crocea head kidney cell line comprises the steps of (1) preparing a cell culture medium; (2) constructing primary culture; (3) performing secondary culture. The constructed pseudosciaena crocea head kidney cell line is fibrous, can be subjected to continuous passage, and is sub-cultured for 70 generations; a large quantity of pseudosciaena crocea head kidney cell lines can be provided; the pseudosciaena crocea head kidney cell line can be directly applied to research of various pseudosciaena crocea immunity related functional genes. The pseudosciaena crocea head kidney cell line can be applied to important researches including cytobiology, molecular biology, virology, toxicology, oncology, genetics, immunology and the like.

The invention discloses a method for characterizing a microorganism phagocytosis function of macrophages of trachinotus ovatus. The trachinotus ovatus is dissected and separated, and the macrophages of the head kidney of the trachinotus ovatus are obtained and are extracted with a Percoll solution density gradient centrifugation method. The number of survived cells cultured in vitro is calculated with a trypan blue staining method, and the survival condition of the cells in one week is detected. The phagocytosis of the macrophages of the head kidney of the trachinotus ovatus on Nocardia seriolea, saccharomycetes and the like is determined through a phagocytosis experiment, so that the phagocytosis function of the macrophages is observed. An experimental result shows that the macrophages of the trachinotus ovatus can be extracted very well, and the survival condition and the phagocytosis condition of the macrophages of the trachinotus ovatus can be observed very well. A foundation is laid for further research of physiology, pathology and toxicology of the macrophages of the trachinotus ovatus.