43 results about "Edwardsiella sp." patented technology

A colloidal gold immunochromatographic test strip for rapid detection of Edwardsiella tarda, the invention relates to a method of using nano-colloidal gold particle-labeled antibody technology and double-antibody sandwich immunochromatography to detect antigen patterns, and prepared a colloid of Edwardsiella tarda Gold immunochromatographic test strips. A colloidal gold immunochromatographic detection test strip for Edwardsiella tarda and its preparation method and application are provided. The test strip contains sample pads, gold standard pads, nitrocellulose membranes with detection line and quality control line antibodies, absorbent pads, and bottom plates. The sample pads, gold standard pads, nitrocellulose membranes, and absorbent pads are sequentially overlapped and connected. Assembled by pasting on the bottom plate. The detection application of the test strip is simple in operation, good in specificity, high in sensitivity, convenient and fast, does not require special equipment and professional training, and the results are clear and easy to distinguish and easy to promote. It is suitable for detection of infection and contamination of Edwardsiella tarda.

The invention relates to the field of molecular biology and immunology and particularly discloses an Edwardsiella tarda recombinant subunit vaccine as well as a preparation method and application thereof. The vaccine has a base sequence in a sequence table SEQ ID No.1. The preparation method of the Edwardsiella tarda recombinant subunit vaccine comprises the following steps of constructing a vaccine antigen expression plasmid pETXV21, converting the vaccine antigen expression plasmid pETXV21 into escherichia coli BL21 (DE3), recovering a recombinant protein after induced expression, mixing the recombinant protein with a strain B187, and dissolving the mixture in PBS (Phosphate Buffer Saline) to obtain a vaccine mixed liquor which has the function of immune protection to Edwardsiella tarda. The vaccine mixed liquor is intraperitoneally injected to fishes to achieve the purpose of immune protection.

The invention relates to the field of immunology, and specifically relates to an Edwardsiella tarda subunit vaccine and a preparation method thereof. The protein gene of the vaccine has a base sequence shown in SEQ ID No.1 in a sequence table, and the vaccine protein coded by the protein gene has an amino acid sequence as shown in SEQ ID No.2 in the sequence table. The subunit vaccine provided by the invention is mixed with an oil emulsion adjuvant, and then capable of effectively exciting the generation of specific antibodies after immunizing the fish through intraperitoneal injection; the Edwardsiella tarda subunit vaccine is capable of efficiently improving the ability of resisting the infection of the Edwardsiella tarda of the immune fish and thus can be applied to controlling the Edwardsiella tarda of the fish.

The invention provides four gene deletion mutants of wild edwardsiella tarda (E.tarda) and derivative strains thereof. The four gene deletion mutants are respectively an E.tarda<deltaEvpC> strain with an EvpC gene losing function and derivative strains thereof, an E.tarda<deltaEvpCdeltaEsrB> strain with an EsrB gene losing function and derivative strains thereof, an E.tarda<deltaEvpCdeltaEsrBdeltaPstB> strain with a PstB gene losing function and derivative strains thereof, and an E.tarda<deltaEvpCdeltaEsrBdeltaPstC> strain with a PstC gene losing function and derivative strains thereof. 1-3 genes such as EvpC, EsrB, PstB, PstC and the like of the attenuated strains have parts causing loss of function or suffer from complete deletion, point mutation, shifting or insertion. Compared with a wild 1101 strain or other virulent strains, the E.tarda<deltaEvpC> strain, the E.tarda<deltaEvpCdeltaEsrB> strain, the E.tarda<deltaEvpCdeltaEsrBdeltaPstB> strain and the E.tarda<deltaEvpCdeltaEsrBdeltaPstC> strain have the advantage that the virulence does not exceed 1/40, 1/400, 1/4000 and 1/4000 of the virulence of the wild E.tarda 1101 strain or other virulent strains. The genetically engineered attenuated strains of E.tarda can be applied as attenuated vaccine strains of E.tarda, genetic engineering vectors or probiotics.

The invention provides a breeding method of excellent strains of hybrid pelteobagrus fulvidraco resistant to Edwardsiella ictaluri.The method comprises the following steps: collecting the pelteobagrusfulvidraco and pelteobagrus vachelli, cultivating till mature, and then reproducing to obtain the fry of purebred pelteobagrus fulvidraco and the pelteobagrus vachelli with different populations; soaking all the fingerlings of the pelteobagrus fulvidraco and the pelteobagrus vachelli with a soaking method according to determined medial lethal concentration of the Edwardsiella ictaluri; performingbreeding on the population resistant to the Edwardsiella ictaluri twice, screening the excellent disease-resistant strains by the survival rate of mixed-cultured fry, respectively cultivating the strains of the pelteobagrus fulvidraco resistant to the Edwardsiella ictaluri, and the pelteobagrus vachelli to sexual maturity, and then producing the hybrid pelteobagrus fulvidraco according to a universal artificial reproduction method of fish. According to the breeding method of the excellent strains of the hybrid pelteobagrus fulvidraco resistant to the Edwardsiella ictaluri, hole-in-head-disease-resisitant parents of the pelteobagrus fulvidraco and the pelteobagrus vachelli can be cultivated, the incidence and mortality of hole-in-head disease in the seedling and culture process are improved, and the survival rate of the culture is improved.

The invention provides turbot-derived Edwardsiella piscicida and an application thereof. The turbot-derived Edwardsiella piscicida is an Edwardsiella piscicida H4-S18 strain, the gene sequences of theEdwardsiella piscicida H4-S18 strain are shown as SEQ ID No.1, SEQ ID No.2 and SEQ ID No.3, and the Edwardsiella piscicida H4-S18 strain is preserved with the preservation number of CCTCC NO: M 2020450. The invention further relates to an application of the turbot-derived Edwardsiella piscicida in preparation of vaccines. The turbot-derived Edwardsiella piscicida provided by the invention is obtained by screening from diseased turbots, is the turbot-derived Edwardsiella piscicida carrying drug resistance genes for the first time in China, the turbot-derived Edwardsiella piscicida carrying thedrug resistance genes is applied for the first time to prepare the vaccines, and therefore, the immune protection rate of the turbots to the drug resistance Edwardsiella piscicida is effectively improved.

The invention relates to the field of molecular biology, in particular to an application of serine protease (PoCFI-Tryp) of a flounder complement regulatory protein I factor. The invention relates tothe application of serine protease (PoCFI-Tryp) in the flounder complement regulatory protein I factor in preparation of a bacteriostatic agent. The serine protease disclosed by the invention can be combined with various bacteria, and can obviously inhibit the growth of Edwardsiella tarda, Vibrio anguillarum, Vibrio harveyi and Streptococcus iniae. The obtained protein has the application potential in bacterial infection resistance.

The invention relates to a microbe detection method and detection kit, particularly a fluorescent probe PCR (polymerase chain reaction) method for simultaneously detecting Edwardsiella tarda and Edwardsiella ictaluri. The method comprises the following step: carrying out real-time fluorescence PCR reaction by using bacteria DNA (deoxyribonucleic acid) as a template by utilizing specific probes and primer pairs. The method is characterized in that the primer primers are respectively sequences disclosed as SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4; the SEQ ID NO.1 is SEQ ID NO.1: 5'-CTGCCAAGCAGGGACGTAA-3', SEQ ID NO.2: 5'-CGAGGAGAGCATCTTGTCGAA-3', SEQ ID NO.3: 5'-TCCATTCGTCTGTGCGACAA-3', and SEQ ID NO.4: 5'-AAAACACGTTCGGATGGATTG-3'; the probes are nucleotide sequences which is identical to or complementary with the sequences between the primer pair on the Edwardsiella tarda or Edwardsiella ictaluri genome DNA; the 5' end and 3' end of the probes are respectively labeled with a fluorescence report group and a fluorescent quenching group; the probes are sequences disclosed as SEQ ID NO.5 and SEQ ID NO.6; and the SEQ ID NO.5 is 5'-ATCCTCAACGTCGAGAAGGCGCG-3', and the SEQ ID NO.6 is 5'-CACTATGAGGGTGGGATCAAGGCGTTT-3'.

The invention provides a kit for rapidly detecting Edwardsiella ictalurus of fishes and a detection method. The kit comprises a slide treated by polylysine, a rabbit anti-Edwardsiella ictalurus polyclonal antibody, an HRP labeled goat anti-rabbit lgG, a confining liquid and a developing liquid. The slide is used as a solid-phase carrier, so that the detection cost is greatly reduced; the rabbit anti-Edwardsiella ictalurus polyclonal antibody prepared by the invention can recognize a plurality of epitopes of the same antigen, and is strong in specificity and high in sensitivity; TMB is used asa peroxidase substrate, TMB is used as a developing solution, a soluble light blue product can be generated, and the light blue product is adsorbed on the filter paper, so that the color change can bevisually observed by naked eyes. The detection method is low in equipment requirement, low in cost, simple and convenient to operate, green, environment-friendly and high in sensitivity.

The invention relates to a novel Edwardsiella attenuated target and application thereof. The gene ETAE-0023 related to the toxicity of the Edwardsiella is identified and obtained in the Edwardsiella for the first time. The invention also discloses an Edwardsiella attenuated strain and an attenuated vaccine which are developed by taking the gene as a target, and the Edwardsiella attenuated strain and the attenuated vaccine have remarkable immunogenicity, high in-vivo clearance rate and very ideal safety and immune response effect.

The invention discloses a body fluid small molecule metabolic marker for zebra fish infected with Edwardsiella tarda. The marker is at least one of arginine, proline, valine, leucine, isoleucine and Cadaverine, preferably proline and valine. The invention also discloses an application of the body fluid small molecule metabolic marker of zebra fish infected with Edwardsiella tarda in improving theimmunity of zebra fish responding to Edwardsiella tarda infection, and an application in regulating and controlling zebra fish arginine proline metabolic pathways and valine leucine isoleucine biosynthetic metabolic pathways. It is found through experiments that proline and valine play an important role when zebra fish responds to Edwardsiella tarda infection, play an extremely important role in zebra fish innate immune response and can be used for research of metabolome functions. The marker has important reference value for researching metabonomics related to zebra fish response to Edwardsiella tarda infection immune response.

The invention relates to a novel edwardsiella attenuated strain, wherein the expression of a target gene or a protein coded by the target gene is regulated and controlled. The edwardsiella attenuated strain can be used as a preventive drug for edwardsiella diseases, and has extremely obvious effect on inhibiting edwardsiella infection.

The invention relates to a hybrid pelteobagrus fulvidraco breeding method for preventing and controlling Edwardsiella diseases, and belongs to the technical field of aquaculture. The method comprisesthe following operation steps of (1) before stocking, carrying out targeted control over parents of whole-pond glochidia, and putting fingerlings of hybrid pelteobagrus fulvidraco into a mixed solution for medicated bath before stocking; (2) fingerling stocking: stocking hybrid pelteobagrus fulvidraco fingerlings, bighead carp fingerlings, silver carp fingerlings and crucian fingerlings in a mixedmanner; (3) feeding special puffed floating granulated feed for pelteobagrus fulvidraco in the whole process; (4) enhancing immunity: from the middle ten days of May to the middle ten days of October, adding an immunopotentiator into the feed in the whole process; (5) water quality substrate improvement: from the middle ten days of May to the middle ten days of October, respectively and regularlyusing microbial preparations such as bacillus, lactic acid bacteria, photosynthetic bacteria and EM bacteria for regulating the water quality; and (6) regularly expelling insects: from June to September, using a protozoa insect repellant once in the morning of sunny days monthly to expel trichodiniasis, chilodoniasis and the like in a targeted manner every month. The morbidity of the hybrid pelteobagrus fulvidraco is reduced to be less than 20%, and the mortality is reduced to be less than 15%.

The invention provides a fish vaccine combined immunization inoculation method. A first mode is characterized in that vaccine preparation and injection are performed according to the antigen concentration of a late pure edwardsiella attenuated candidate vaccine strain WED and a turbot vibrio anguillarum genetic engineering live vaccine MVAV6203 according to the ratio of 1: 10 to inoculate turbots; and a second mode is characterized in that the turbot in the fry stage is selected, and is soaked in and inoculated with the lagged pure edwardsiella attenuated candidate vaccine strain WED strain, and the turbot vibrio anguillarum genetic engineering live vaccine MVAV6203 strain is inoculated by injection when the turbot grows to the juvenile fish stage. According to the invention, a cold water flounder culture variety is used as an immune target animal, a combined inoculation strategy of the two vaccines is explored, and the feasibility and effectiveness of the combined inoculation strategy are determined by evaluating the influence of the strategy on key production performances such as feed conversion rate, death and culling rate and the like of flounder in actual production. Finally, a novel flatfish healthy breeding production system with combined immunization as the core is established, and the combined prevention and combined control effect on multi-pathogen mixed infection is achieved.

The invention discloses an eel Vibrio vulnificus/Edwardsiella tarda duplex recombinant protein and a preparation method thereof. The duplex recombinant protein comprises a Vibrio vulnificus outer membrane protein OmpU extramembrane part and an Edwardsiella tarda outer membrane protein OmpA extramembrane part which are connected together, wherein the Vibrio vulnificus outer membrane protein OmpU extramembrane part and Edwardsiella tarda outer membrane protein OmpA extramembrane part respectively comprise 238th-456th amino acid sequences and 467th-695th amino acid sequences in SEQ ID NO.01. The Vibrio vulnificus outer membrane protein OmpU extramembrane part and Edwardsiella tarda outer membrane protein OmpA extramembrane part are connected together by a gene engineering means to establish the duplex recombinant protein. After being used for immunizing the eel, the protein can simultaneously resist the infection of Vibrio vulnificus and Edwardsiella tarda. Compared with the duplex inactivated vaccine, the duplex recombinant protein has higher relative immunity protection effect and lower toxic and side effects.

The invention discloses an edwardsiella catfish bacteriophage EIP-1 and application thereof, and belongs to the technical field of biology. The invention discloses a strain of edwardsiella catfish bacteriophage EIP-1, the preservation number of which is CGMCC (China General Microbiological Culture Collection Center) No.45077. The separation and identification of the bacteriophage not only enriches a bacteriophage germplasm resource library, but also provides a selectable potential scheme for solving the problem of fish death caused by Edwardsiella catfish infection in aquaculture; through whole genome sequencing of the bacteriophage EIP-1, functional genes and functional proteins of the bacteriophage EIP-1 can be further deeply excavated, and one or more enzymes with a wide-spectrum splitting effect are provided for bacteriophage treatment and biological prevention and treatment of Edwardsiella catfish infection in aquaculture, so that the application universality and effectiveness of the bacteriophage are improved; the bacteriophage EIP-1 can effectively split edwardsiella catfish and is used for preventing and treating fish infection death caused by edwardsiella catfish infection in aquaculture.