80 results about "Edwardsiella" patented technology

The invention relates to an Edwardsiella tarda strain and an application method thereof. The Edwardsiella tarda strain is separated from a turbot adult fish body and is a wild strain with strong virulence, and the preservation number of the Edwardsiella tarda strain is CGMCC No.7197. Preparation modes of an antigen of the Edwardsiella tarda strain comprise any one or more than one of an inactivated thallus, a bacteruak ghost ingredient, an attenuated strain, a protective antigen, an antigen subunit and an expression product of an antigen determinant or an antigen gene expression carrier; the produced vaccine can be a single ingredient of the antigen prepared by utilizing the Edwardsiella tarda strain and can also be a combined vaccine produced by mixing the antigen prepared by utilizing the Edwardsiella tarda strain with antigens of other bacteria, and the prepared single or combined vaccine antigen is added with an adjuvant to produce the vaccine; and an inoculation mode of the vaccine in immunization application can adopt injection immunization, wound immunization, immersion bath immunization or oral administration immunization.

The invention relates to the field of fish medicine preparations, in particular to Sanhuang supper micropowder as well as a preparing method and applications thereof. The supper micropowder comprises in percentage by weight as follows: 25-35% of baical skullcap roots, 25-35% of amur cork tree bark, 25-35% of rhubarb and 5-10% of indigowoad leaves. The Sanhuang supper micropowder has effects of clearing heat, cooling blood, resisting viruses and detoxifying, plays powerful antibacterial, disinfection and antiviral roles in water bodies and pathogenic bacteria of aqueous animal (gills, body surfaces and the like) tissue and is particularly used for mainly treating bacterial enteritis caused by pseudomonas, vibrio, aeromonas hydrophila and Edwardsiella in the aqueous animals. Tests prove that the Sanhuang wall-breaking supper micropowder can improve and treat white excrement enteritis of penaeus vannamei boone and septicemia of Micropterus salmoides to a certain extent, can enhance the aeromonas hydrophlia infection resistance of the Micropterus salmoides and is a novel fish medicine for effectively protecting intestinal tracts and preventing bacterial enteritis in the aqueous animals, and the use amount can be significantly decreased by 2-3 times.

The invention relates to an adjuvant for improving the immunization effect of Edwardsiella vaccine and a use method of the adjuvant. The adjuvant is characterized by being extracted from raw materials including grain, yeast and a part of fungi or algae. The effective component of the adjuvant is beta-1,3-glucan or the natural, artificially modified or synthetic product of the beta-1,3-glucan. The adjuvant can be used for increasing the specific immune protection ratio of any one or more inculcated Edwardsiella vaccine antigens including the inactivated bacteria, the bacteria disintegration component, the less-virulent strain, the attenuated strain, the protective antigen, the antigen subunit, the antigenic determinant clusters or the expression product of the antigen cell expression vector of the Edwardsiella, can be used together or not together with the vaccine antigen and can be prepared into a single preparation which is used together with the vaccine antigen.

The present invention encompasses a recombinant Edwardsiella bacterium, and compositions and methods of use thereof.

The present invention discloses a technique for preventing and curing the diseases of aquacultural animals, which relates to an attenuated vaccine for aquacultural bacterial diseases, in particular to an attenuated live vaccine for Edwardsiella tarda. The esrB gene of Edwardsiella tarda strain LSE40 loses an attenuated mutant strain, the Edwardsiella tarda mutant strain is MZLSE40esrB, which is preserved in China General Microbiological Culture Collection Center (CGMCC) and the accession number of which is CGMCC No.2087, and the mutant strain MZLSE40esrB does not contain exogenous antibiotic selectable markers and exogenous gene fragments. In respect to the wide type Edwardsiella tarda, the attenuated live vaccine has obviously low toxicity and immune protective rate and does not contain any exogenous antibiotic resistance marker and exogenous gene fragment. Experiments show that the attenuated live vaccine can effectively protect susceptible fishes from being infected with the pathogenic Edwardsiella tarda.

The present invention encompasses a recombinant Edwardsiella bacterium, and compositions and methods of use thereof.

The invention relates to the field of biotechnology, in particular to primer sequences for detecting Edwardsiella tarda, Edwardsiella ictarda or pathogenic Edwardsiella tarda, and a detection method and application thereof. In the method for polymerase chain reaction (PCR) detection through primers, primers for detecting pathogenic and nonpathogenic Edwardsiella and a method for single or multiplex amplification detection by applying the primers are further related; and the invention further relates to a kit comprising the primers and used for detecting the Edwardsiella tarda, Edwardsiella ictarda or pathogenic Edwardsiella tarda. By adopting multiplex PCR amplification, more than two pairs of primers are present in the same reaction system and can amplify more than two target genes, so that the multiplex PCR is reagent-saving, time-saving and labor-saving than single PCR; therefore, the cost is reduced and the efficiency is improved.

The invention discloses eel aeromonas hydrophila and edwardsiella tarda bigeminal recombinant protein and a preparation method thereof. The bigeminal recombinant protein disclosed by the invention comprises an aeromonas hydrophila type II pore protein membrane outer part and an edwardsiella tarda ompS2 membrane outer part which are connected together, and the amino acid sequence of the bigeminal recombinant protein is as shown by SEQ ID NO1. According to the preparation method disclosed by the invention, the aeromonas hydrophila type II pore protein membrane outer part and the edwardsiella tarda ompS2 membrane outer part are connected together through a genetic engineering means to construct the bigeminal recombinant protein, and the bigeminal recombinant protein can prevent inflections of aeromonas hydrophila and edwardsiella tarda at the same time after being used for immunizing eels, and has a better immune effect compared with immunization with single outer membrane protein. The bigeminal recombinant protein disclosed by the invention can obtain an immune effect on a variety of pathogenic bacteria through only once injection on a fish body, so that injuries to the fish body are reduced, and immunization steps are simplified; the preparation method of the bigeminal recombinant protein is suitable for industrial production.

The invention relates to an edwardsiella tarda subunit oral microencapsule vaccine for an aquatic product. The edwardsiella tarda subunit oral microencapsule vaccine comprises a calcium alginate-chitosan release-controlled microcapsule and three recombinant protein-lipopolysaccharide conjugates which are encapsulated in the microcapsule, wherein the three recombinant protein-lipopolysaccharide conjugates include edwardsiella tarda glyceraldehyde-3-phosphate dehydrogenases, outer membrane protein and flagellin which are mixed with astragalus polysaccharide. The oral microencapsule is characterized in that the encapsulation efficiency is 78%, the protein load is 102mg / g, and the grain size is 150+ / -10 microns. The prepared oral microencapsule vaccine is convenient to use, proper in grain size, high in slow release performance and high in antigen protection performance, and can effectively irritate a fish intestinal immune system to perform immune response so as to enable a fish to gain the specific immunity protection capacity for edwardsiella tarda; and as a result, the edwardsiella tarda subunit oral microencapsule vaccine is applicable to prevention and treatment of fish edwardsiella tarda disease.

The invention discloses a traditional Chinese medicine preparation for treating the Chinese soft-shell turtle rotten-skin disease caused by edwardsiella and a preparing method of the traditional Chinese medicine preparation. The traditional Chinese medicine preparation is medicine powder prepared from, by weight, 15-25% of isatis root, 5-15% of rheum officinale, 5-15% of sophora flavescens, 5-15% of humifuse euphorbia herb, 5-15% of Chinese gall, 5-13% of liquorice, 5-13% of folium isatidis, 5-15% of amur cork tree bark, 5-13% of houttuynia cordata and 3-9% of fructus forsythiae. The sum of the percentages of all the components is 100%. The traditional Chinese medicine preparation for treating the Chinese soft-shell turtle rotten-skin disease caused by the edwardsiella has the advantages of being good in treating effect on treating the Chinese soft-shell turtle rotten-skin disease caused by the edwardsiella and free of drug residues.

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.

Disclosed is a vaccine for edwardsiellosis and streptococcosis in a fish. Specifically, disclosed is a vaccine for edwardsiellosis in a fish, which comprises inactivated cells of (A) an Edwardsiella tarda strain derived from the target fish, and inactivated cells of (B) an Edwardsiella tarda strain derived from a fish other than the target fish, wherein, when the strain (A) is a typical Edwardsiella tarda strain, the strain (B) is an atypical Edwardsiella tarda strain, whereas when the strain (A) is an atypical Edwardsiella tarda strain, the strain (B) is a typical Edwardsiella tarda strain. Also specifically disclosed is a vaccine for edwardsiellosis and / or streptococcosis in a fish, which comprises the components mentioned above and inactivated cells of (C) Streptococcus iniae and / or Streptococcus parauberis.

The invention provides an Edwardsiella tarda immunogenic protective antigen which is an Edwardsiella tarda flagellar related protein with an amino acid sequence expressed in SEQ ID NO:2 and a preferred nucleotide sequence expressed in SEQ ID NO:1, a recombination expression vector corresponding to the Edwardsiella tarda immunogenic protective antigen, an Edwardsiella tarda subunit vaccine prepared by the Edwardsiella tarda immunogenic protective antigen, and an application of the Edwardsiella tarda immunogenic protective antigen in the preparation of the Edwardsiella tarda subunit vaccine. The Edwardsiella tarda immunogenic protective antigen provided by the invention has a good immunogenicity and can be used for preparing a safe, effective and economic vaccine, thus has a high commercialvalue and is suitable for large scale popularization and application.

The invention discloses a method for breeding Edwardsiella tarda-resistant excellent flounder breed. The method mainly comprises the following steps: screening Edwardsiella tarda-resistant pedigrees specific microsatellite markers by an Edwardsiella tarda-infected flounder fry and disease resistance measuring method, an Edwardsiella tarda-resistant group screening method, an Edwardsiella tarda-resistant pedigrees screening method, an Edwardsiella tarda-resistant pedigrees specific microsatellite marker screening method and the like; establishing a disease-resistant pedigree offspring disease resistance and breeding survival rate measuring method, and establishing the Edwardsiella tarda-resistant excellent flounder breed. The flounder breed bred by the method has the characteristics of strong Edwardsiella tarda resistance, high breeding survival rate, low death rate and relatively high growth speed; the problem of massive death caused by an ascites disease due to the infection of Edwardsiella tarda in summer is basically solved; the method is suitable for being popularized and applied in flounder breeding farms in coastal areas in China.

The invention relates to an application of nonpathogenic edwarsiella tarda, belonging to the fields of immunology and microbiology. The nonpathogenic edwarsiella tarda has immune and protective effect on the infection of pathogenic edwarsiella tarda and high protective action on the pathogenic edwarsiella tarda and also has easy preparation and immune operation without any processing process.

The invention discloses a preparation capable of improving sensibility of bacteria on antibiotics and relates to glycine and glucose. Because glycine serving as small molecules capable of improving the sensibility of bacteria on antibiotics achieves an effect of inhibiting drug-resistance bacteria, glycine can be applied to improve the sensibility of bacteria on antibiotics. Because glycine and glucose serving as small molecules capable of improving the sensibility of bacteria on antibiotics achieve an effect of inhibiting drug-resistance bacteria, glycine and glucose can be combined to be applied to improve the sensibility of bacteria on antibiotics. The preparation is composed of glycine, glucose and antibiotics in a mass ratio of 1:(0.01-100):(0.01-10). A usage amount of glycine is 0.1mg-1g / kg per time. After glycine is added, the survival rate of slow Edwardsiella drug-resistance bacteria treated by kanamycin is obviously decreased, meaning the sensibility of slow Edwardsiella drug-resistance bacteria to kanamycin can be improved.

The invention discloses a preparation method of astragalus polysaccharide chitosan nanoparticles. The astragalus polysaccharide chitosan nanoparticles prepared by the method have good prevention and treatment effects on Edwardsiella catarrhalis. According to the preparation method, the astragalus polysaccharide chitosan nanoparticles are prepared by applying an ionic crosslinking method for the first time; the degradation of the medicine before reaching an acting part is reduced, the release speed of the medicine is reduced, the release time is prolonged, and the stability and in-vivo availability of astragalus polysaccharide are improved, so that the curative effect is improved, the administration frequency and dosage are reduced, and the product is more conveniently applied to fish culture.

The invention discloses gene amplification primers for quickly detecting Edwardsiella trada. Specific primers are designed by taking esrB of E. tarda or a hlyB upstream sequence of the Edwardsiella trada as a target. A gene amplification detection method by the primers has the characteristics of simplicity, convenience, quickness, specificity and sensitivity; genes with 10 copy number can be detected in 40 minutes only through a water bath or a metal bath; and the method is suitable for quickly detecting the E. tarda in field and has good application prospect.

The invention provides a preparation method of channel catfish Edwardsiellosis vaccines. The preparation method includes the steps of strain extraction, infection, material acquiring, suspension preparation, enlarged cultivation, counting, inactivation, synthesizing, diluting, split charging and inspection. The channel catfish Edwardsiellosis vaccines are prepared by adding a proper amount of double-resistant and antiseptic agent into Edwardsiella. The preparation method is low in cost and convenient in operation, immunity of channel catfishes can be improved effectively, and channel catfish Edwardsiellosis is prevented.

The present invention relates to a field of vaccinology, and especially relates to a bacteria-delivered cross protection vaccine against Edwardsiella tarda and Streptococcus iniae. The cross protection vaccine is a recombinant strain obtained by plasmid [rho]DKS10 transforming escherichia coli DH5[alpha]. The vaccine obtained in the present invention has 83% and 75% protection efficiency respectively for Edwardsiella tarda and Streptococcus iniae infection.

The invention discloses a Weissella confusa 3DW having a high antagonism effect on Edwardsiella tarda. The Weissella confusa 3DW is preserved in China General Microbiological Culture Collection Centerwith the preservation codename being CGMCC 14656. A difference between the inhibition zone diameter and the colony diameter of the Weissella confusa 3DW on the Edwardsiella tarda is 8.17 mm, so the Weissella confusa 3DW can effectively prevent and treat various infected diseases of aquatic products, caused by the Edwardsiella tarda. The strain can significantly reduce the pH value of water, can be applied to a micro-ecologic preparation to reduce the pH value of water in a saline and alkaline land culture pond, and is suitable for saline and alkaline land pond culture.

The invention relates to the field of aquaculture, and discloses lactobacillus acidilactici, an aquatic feed additive based on the lactobacillus acidilactici, fish feed and application of the lactobacillus acidilactici, the aquatic feed additive and the fish feed. The lactobacillus acidilactici can promote the growth performance of cultured fishes, improve the immunity of the cultured fishes, especially improve the capacity of resisting Edwardsiella infection of longsnout catfish, well adapt to the digestive tract environment of the cultured fishes and stably play a role.

The invention provides a composite bacteria preparation used for disinfection. The composite bacteria preparation is composed of, by mass, 1 to 25% of a microbial community, 0.5 to 10% of a microbial nutrient solution, and 65 to 95% of bacteria normal saline. The microbial community with bacteria killing effect is taken as a main microbial community, the content of effective probiotics is capable of reaching 3*10<7> cfu / g; the main microbial community possesses killing, phagocytosis, and rapid lysis effect on pathological bacteria such as vibrio parahaemolyticus, aeromonas hydrophila, aeromonas punctata, escherichia coli, salmonella, and edwardsiella spp, so that the composite bacteria preparation is capable of reducing the content of pathological bacteria in water bodies obviously, killing bacteria with bacteria, and repairing water body microecological balance.

The invention provides a genetic engineering vaccine for an anti-idiotypic antibody of edwardsiella tarda and a method for preparing the same. The method comprises the following steps: adopting an RT-PCR method to clone VH and VL variable region genes from a hybridoma cell strain which excretes idiotype monoclonal antibodies of the edwardsiella tarda; introducing a connecting peptide (Gly4Ser)3 into the VH and VL variable region genes; constructing a single chain antibody scFv gene in the form of VL-Linker-VH in vitro, and cloning the single chain antibody scFv gene to an expression vector PET28a and expressing the same in Escherichia coli; and after purifying expressed product, identifying the product through SDS-PAGE, Western blot and ELASA: expressing an anti-idiotypic single chain antibody gene scFv of the edwardsiella trada in a BL21 bacterium, preserving the expressed product in the form of an insoluble inclusion body, obtaining a high purity single chain antibody fragment by dissolving the inclusion body, purifying, and renaturing in vitro. The relative molecular weight of the recombinant protein is 27 kD. The immune protection rate for sea fishes is averagely more than 80 percent.

The invention provides a preparation method of eel edwardsiella phage, a microecological preparation and application, and belongs to the technical field of eel culture. The preparation method of the eel edwardsiella phage comprises the following steps that 1, a liquid culture medium is inoculated with eel edwardsiella tarda, fermentation culture is carried out, and fermented liquid is obtained; 2,the phage of the eel edwardsiella phage is directly added to the fermented liquid, and a phage culture system is obtained for phage culture; 3, when phage culture is carried out for 10 h to 20 h, edwardsiella liquid starts to flow into the phage culture system, wherein the concentration of edwardsiella in the phage culture system ranges from 108 CFU / ml to 1014 CFU / ml; 4, when phage culture is finished 18 h to 24 h later, and the eel edwardsiella phage is obtained. The method is easy to operate and short in culture period, and the obtained phage is high in potency.

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 the field of molecular biotechnology, in particular to a rapid identification method of Edwardsiella fisicida sRNA. The method provided by the invention comprises the following steps: constructing an Edwardsiella fisicida hfq gene deletion mutant strain; the expression levels of sRNA in wild strain and hfq mutant were detected by fluorescence quantitative PCR, and the dependency of sRNA on Hfq protein was determined according to the difference of expression levels between wild strain and hfq mutant. The method provided by the invention can quickly and accurately determine the dependency relationship between the sRNA and the Hfq protein, has simple operation steps, does not need the high technical difficulty experiment operation such as RNA immunoprecipitation and the like, can realize the high-throughput detection and identification, greatly reduces the detection cost, and is easy to be widely applied.

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 relates to the field of vaccinology, in particular to a cross protection vaccine specific to Edwardsiella tarda and Vibrio harveyi. The cross protection vaccine is a recombination bacterial strain that recombines and converts plasmid pAQ1 to Edwardsiella tarda ATCC15947. The cross protection vaccine is a cross protection vaccine of Edwardsiella tarda and Vibrio harveyi. The vaccine has over 90% protective effect on infection of Edwardsiella tarda and Vibrio harveyi.

A live attenuated Edwardsiella ictaluri bacterium lacking a viable gene encoding a functional evpB protein and a method of using the same to protect fish against infection from virulent Edwardsiella ictaluri. The methods and compositions for protecting fish against infection from virulent Edwardsiella ictaluri comprising administering to a fish a therapeutically effective amount of an attenuated Edwardsiella ictaluri bacterium lacking a viable gene encoding a functional EvpB protein. The bacterium may include an insertion and / or deletion mutation in the evpB gene. The fish include catfish, preferably catfish fingerling or a catfish fry. The composition may be delivered via immersion delivery, an injection delivery, an oral delivery, or combinations thereof.