135 results about "Vibrio anguillarum" patented technology

The present invention discloses bacillus amyloliquefaciens and application thereof in aquaculture. The bacillus amyloliquefaciens SIP0902 strain is preserved in the China Center For Type Culture Collection (CCTCC), AND the preservation number is CCTCC NO:M2014323. The bacillus amyloliquefaciens is separated from intestinal tract of penaeus vanmamei, is an endogenous strain, and has strong antagonism to vibrio parahaemolyticus, vibrio anguillarum, vibrio alginolyticus and other 7 kinds of aquatic animal pathogenic vibrios. Bacillus amyloliquefaciens powder prepared from the bacillus amyloliquefaciens can significantly enhance non-characteristic immune function of the penaeus vanmamei, and can improve the pathogen infection resisting ability of prawn.

The present invention relates to a detection chip for performing gene detection to various vibrio and its detection and applications. The invention provides 16S rRNA sequences corresponding to each vibrio of vibrio anguillarum, vibrio harveyi, vibrio alginolyticus, vibrio parahaemolyticus, brilliant vibrio and Fisher vibrio; heat shock protein hsp60 probe sequence; virulence gene probe sequence; 16S rRNA forward primer sequence; 16S rRNA reverse primer sequence; heat shock protein hsp60 forward primer sequence; heat shock protein hsp60 reverse primer sequence; virulence gene forward primer sequence and virulence gene reverse primer sequence. The present invention has specific, sensitive and high-throughput features, can simultaneously detect six kinds of bacteria virulence genes, and the invention will effectively guide the production as an important disease early-warning detection method used in clinical diagnosis of aquatic animals.

The invention relates to a selective breeding method for disease-resistant superior strains of paralichthys olivaceus, which comprises group selective breeding and family selective breeding of the paralichthys olivaceus and is characterized by also comprising molecular marker-assisted selective breeding, manual gynogenesis selective breeding or inbreeding selective breeding of a disease-resistant paralichthys olivaceus family. Multiple kinds of biotechnology such as group selective breeding, intraspecific hybridization, family selective breeding, molecular marker-assisted selective breeding, manual gynogenesis selective breeding, inbreeding and the like is comprehensively adopted; a selective breeding method for the disease-resistant superior strains of marine fishes in China is established for the first time by taking the paralichthys olivaceus as a material; the superior strains of the paralichthys olivaceus of which the vibrio anguillarum resistance is obviously improved are bred; and the breeding survival rate is improved by 20 to 47 percent and the growth rate is improved by over 20 percent. The method opens up a new technical approach for the selective breeding of the disease-resistant strains of the fishes, is suitable for popularization and application to all cultured fishes, has great significance and application value for breeding the disease-resistant superior strains of the cultured fishes, and can generate great economic and social benefits.

The invention discloses a gene chip of aquatic product cultivation pathogenic bacterium, comprising a solid phase carrier which is modified chemically, a detection probe and a quality control probe are distributed on the solid phase carrier in a dot matrix way; the detection probe comprises specificity 16S rDNA sequences and/or gyrB gene sequences of vibrio, comma bacillus, vibrio harveyi, vibrio alginolyicus, vibrio anguillarum, vibrio parahemolyticus, nocardia, nacardia seriolea, aeromonas, hydrophilic aeromonas, streptococcus and dolphin streptococcus, which are to be detected, the quality control probe includes PCR positive, chip fixed positive control, chip hybridizing negative control, chip hybridizing positive control and chip hybridizing blank control; the gene chip has the advantages of small volume and high flux, can detect known and unknown germs of the vibrio, the nocardia, the aeromonas and the streptococcus, and can detect specific germs with multiple kinds, and the simpleness and rapidness and specificity of the germs can be detected, and automatic detection can be carried out after detection software is additionally arranged.

An isolated strain of bacteriophage, specific against bacteria belonging to the Vibrio genre, particularly the anguillarum species, deposited on 3 Oct. 2012 at the Polish Collection of Microorganisms (PCM) of the Ludwik Hirszfeld Institute of Immunology and Experimental Therapy of the Polish Academy of Sciences, with access number F/00072, characterized in that said strain is efficient in the prophylaxis, control and/or treatment of the infection caused by Vibrio anguillarum in all types of species of fish, mollusks and crustaceans that are important for aquaculture susceptible to this bacteria, genome size 48.6 Kb, it is not sensitive to chloroform and its storage temperature is −80° C.

The invention discloses a prawn feed additive, a preparation method thereof, and a composite feed containing the additive. The additive comprises the following raw materials: astragalus, rheum officinale, Codonopsis pilosula, isatis root, humifuse euphorbia herb and gallnut. The additive may also comprise a synergist. The preparation method comprises: weighing various raw materials according to weight percentage; crushing the raw materials; and feeding the various raw materials into a mixer in turn, and uniformly mixing. The composite feed containing the feed, wherein the weight percentage of the additive is 1 to 3 percent. According to the technical scheme provided by the embodiment of the invention: the additive is made according to a formula of Chinese herb medicine, avoids arousing medicine resistance and producing toxic and side effect, prevents medicine residue in prawns, can obviously increase the growth speed of the prawns and improve the immune activity and disease resistance of the prawns, has high resistance for vibrio anguillarum which is pathogenic bacterium, protects the prawns against diseases, greatly improves survival rate and obviously improves production benefit.

The invention relates to a flounders quintuplet inactivated vaccine and a preparation method thereof. The main constituents of the vaccine are mixed by the inactivated bacteria of five bacteria such as the vibrio anguillarum, turbot vibrio, the vibro harveyi, the algae dissolving glue vibrio and the edwardsiella tarda. The preparation method comprises the steps of: respectively preparing the five bacteria into 109-1010cfu/mL of vibrio suspension liquid, inactivating the vibrio suspension liquid by formalin, performing the isovolumetric mixing according to the proportion of 1:1:1:1:1 to prepare the flounders quintuplet inactivated vaccine raw liquid, and evenly mixing the flounders quintuplet inactivated vaccine raw liquid with the 15-35mg/mL astragalus polysaccharides adjuvant raw liquid according to the proportion of 1:1. The flounders quintuplet inactivated vaccine is multiple in immunity effects, can be used for effectively preventing the infection of the five bacterial pathogenic bacteria after being vaccinated for once, and provides a basis for the development of the marine fish multi-germinal inactivated vaccine.

The invention discloses a gene chip for detecting ten types of pathogenic bacteria in sea areas. The gene chip comprises a solid phase slide on which a quality control probe and a detection probe are arranged, wherein the nucleotide sequence components of the detection probe are shown in SEQ ID NO.1-SEQ ID NO.27. The gene chip is capable of simultaneously detecting enterobacter cloacae, edwardsiella tarda, streptococcus faecalis, vibrio fortis, vibrio harveyi, vibrio parahaemolyticus, vibrio splendidus, vibrio vulnificus, vibrio anguillarum and shewanella smarisflavi just in the presence of DNA (deoxyribose Nucleic Acid) without need of live bacteria. The gene chip has remarkable advantages of high throughput, parallelism, miniaturization, automation, rapidness, sensitiveness, quantification, small use amount, and the like.

The invention relates to a method for constructing tongue sole families and breeding good families, which comprises the technical content of breeding a tongue sole group, constructing the tongue sole families and screening a disease-resistant family and a fast growing family of tongue sole. The method comprises the following steps of: constructing the families through the group breeding and intraspecific hybridization technology; then, screening out the family resisting vibrio anguillarum disease from a large amount of family by an artificial infection approach, and screening out the fast growing family by growth comparison. The fries in the good families can be directly popularized and applied, or used for breeding a high-yield and disease-resistant good strain of tongue sole. The invention has the characteristics of strong pertinence, high breeding efficiency, obvious effect and the like. The established method for screening good tongue sole families provides a new technical approach for breeding the high-yield and disease-resistant good strain of tongue sole. The method is suitably popularized and applied to other cultured fishes, and has important significance and application value for breeding high-yield and disease-resistant strains of the cultured fishes.

A Silicibacter sp. strain useful for genetic transformation of marine algae and production of antibiotic agents is described. Silicibacter sp. TM1040 is a genetically tractable member of the marine Roseobacter clade of bacteria that forms an intimate and obligate interaction with algae, and is useful as a probiotic for producing antibacterial agents that are cidally effective against pathogenic bacteria such as Mycobacterium marinum, Vibrio anguillarum, V. coralliilyticus, and V. shiloi bacteria.

The invention discloses a traditional Chinese medicinal recipe for inhibiting vibrio anguillarum. The recipe is characterized by comprising at least one of fructus schisandrae chinensis, pericarpium granati, folium artemisiae argyi, aloe, aluminis usti, catechu, siegesbeckia pubescens mak, radix sanguisorbae, herba lobeliae chinensis, folium ilex, semen arecae, chaenomeles speciosa, sapium sebiferum, moxibustion myrrha, anemone chinensis, herba artemisiae scopariae, rhizoma arisaematis, radix scutellariae, andrographis paniculata, rhizoma polygoni cuspidate slice, fructus quisqualis, herba taraxaci, hops, herba portulacae, fructus forsythiae, gentiana scabra bunge, rhizoma atractylodis, cortex moutan radicis, semen pharbitidis, flos caryophyllata, herba violae, radix clematidis, solanum nigrum, galla chinensis, leguminosae, herba hedyotis, bupleurum chinense, tetradium ruticarpum, fructus mume, folium isatidis, plantago asiatica L and rhizoma curcumae, wherein a recipe which is formed by mixing the pericarpium granati, the folium artemisiae argyi, the fructus schisandrae chinensis and the aloe according to the mass ratio of 1:4:1:4 has an optimal antibacterial effect. The recipe has the advantages of naturality, no toxity and residues, no tolerance and low price.

The invention discloses a method for preparing questin by utilizing an ocean aspergillus flavipes HN4-13 bacterial strain. The method comprises the following steps: conducting separation and purification on antibacterial active substances generated through fermentation of the aspergillus flavipes HN4-13 bacterial strain according to silica-gel column chromatography, Sephadex LH-20 column chromatography, preparation of high performance liquid chromatography (PHPLC) and the like by taking vibrio harveyi as an indicator bacterium, so as to obtain a vibrio harveyi-preventing active compound HY2; indentifying the structure of the active compound HY2 according to spectra data such as ESI-MS, 1H-NMR and 13C-NMR, so as to determine that the active compound HY2 is questin. The questin is obtained through separation of a metabolic product of aspergillus flavipes HN4-13; experiments show that the questin can perform a certain inhibiting function of the pathogenic bacteria, such as vibrio harveyi, vibrio anguillarum, vibrio parahaemolyticus and vibrio cholerae, of aquatic products, and can be used for preparation of medicine preventing pathogenic vibrio of the aquatic products.

The invention relates to a quinones derivative as well as a preparation method of the quinones derivative and an application of the quinones derivative as an antibacterial agent. During the preparation, marine fungi Nigrospora sp.(HM565952) is subjected to strain culture in a culture medium, then, the marine fungi are subjected to fermentation culture in a fermentation culture medium, next, the obtained fermentation liquid is filtered, thalli are removed, and the fermentation liquid is extracted by ethyl acetate; after the extraction liquid is concentrated, chromatographic separation is carried out, and the obtained eluent is concentrated to obtain two red crystals which are respectively 4-Deoxybostrycin and Bostrycin; and acetyl oxide is added into solution in which the two compounds dissolve for reaction to obtain 3-Acetoxy-4-deoxybostrycin and 3-Acetoxy-bostrycin. The quinones derivative provided by the invention has the obvious antibacterial activity on Bacillus subtilis, Bacillus cereus, Micrococcus tetragenus, Micrococcus lutea, Staphylococcus aureus, Staphylococcus albus, Escherichia coli, Vibrio parahaemolyticus and Vibrio anguillarum and can be used for developing antibacterial agents, in addition, raw materials can be produced in large scales, and the resource limitation is avoided, so the application prospect is wide.

The invention discloses four sets of oligonucleotide sequences for recognition and identification of vibrio anguillarum, and a screening method of the oligonucleotide sequences, and relates to the technical field of recognition detection of vibrio anguillarum. Two ends of each oligonucleotide sequence are fixed sequences, intermediate sequences are arrayed according to the order of 5'-3' as follows: a first set contains two sequences of H1 and H2, a second set contains five sequences of H5, H6, H26, H38 and H33, a third set contains three sequences of H12, H25 and H42, and a fourth set contains a sequence of H28; and any of the oligonucleotide sequences in each set serves as the intermediate sequence of an aptamer to be used for vibrio anguillarum detection. The screening method of the sequences sequentially comprises the steps of random oligonucleotide library synthesis, combination, separation, PCR amplification and high-throughput sequencing which are circulated for 3-5 rounds, andfinally, the target sequences are obtained. The sequence length of the aptamer of the vibrio anguillarum is shortened, and the screening method is quick.

The invention provides an anthraquinone derivative, as well as a preparation method and application of the anthraquinone derivative serving as an antibacterial agent. The preparation method comprises the following steps: culturing a strain of Nigrospora sp. (HM565952) in a strain culture medium; fermenting the nigrospora sp. in a fermentation culture medium; filtering the fermentation broth, removing thallus, and extracting the fermentation broth with ethyl acetate; concentrating the extract, and performing chromatographic separation, and concentrating the eluent to obtain two types of yellow powder, namely 3,5,8-Trihydroxy-7-methoxy-2-methylanthracene-9,10-dione and austrocortirubin; and adding an acetic anhydride reagent in a solution dissolved with the compound, and reacting to obtain the anthraquinone derivative. The anthraquinone derivative obtained from the Nigrospora sp. (HM565952) has selective inhibitory activity to Bacillus subtilis, Bacillus cereus, Micrococcus tetragenus, Micrococcus lutea, Escherichia coli, Staphylococcus aureus, Staphylococcus albus, Vibrio parahaemolyticus and Vibrio anguillarum, and can be used for developing selective bacteriostatic agents; and the raw materials can be produced on a large scale, and not limited by resources. Therefore, the anthraquinone derivative has the advantage of wide application prospect.

The invention relates to the technical field of function microorganism screening and particularly provides novel bacillus subtilis SY12 with the preservation number being CCTCC No: M2015483. The bacillus subtilis SY12 has the advantages that the bacillus subtilis SY12 is capable of effectively inhibiting pathogenic bacteria such as vibrio harveyi and vibrio anguillarum, improving immunity of aquaculture animals and reducing diseases; the bacillus subtilis SY12 with high proteinase-producing and amylase producing capabilities can serve as a feed additive, so that digestive ability of the aquaculture animals is improved remarkably, feed use ratio is increased remarkably, and animal growth is promoted.

The invention belongs to the technical field of research and development of medicinal products and relates to a compound lightyellow sophora root preparation and application thereof. The compound lightyellow sophora root preparation is prepared by the following steps of: adding ethanol into lightyellow sophora root, melia azedarach linn leaf and salix leaf respectively; oscillating and lixiviating by using a table concentrator; centrifuging by using a centrifugal machine to obtain supernate; extracting repeatedly for twice; combining the supernate of the three times; filtering; concentrating the filtrate under reduced pressure; drying to obtain extracts of the lightyellow sophora root, the melia azedarach linn leaf and the salix leaf; adding pounded garlic into a round-bottomed flask to perform steam distillation to obtain garlic essential oil; and preparing the compound lightyellow sophora root preparation from the lightyellow sophora root extract, the melia azedarach linn leaf extract, the salix leaf extract and the garlic essential oil. The compound lightyellow sophora root preparation has bacteriostasis on vibrio anguillarum, vibro harveyi, vibrio alginnolyficus and halophilicvibrio. The corresponding preparation can be developed to be used for preventing and treating vibriosis in the aquaculture process. The compound lightyellow sophora root preparation is suitable for industrialized production.

The invention discloses a sinonovacula constricta C1q (Complement 1q) gene, an encoded protein, a cloning method of the sinonovacula constricta C1q gene and a recombinant sinonovacula constricta C1q gene engineering bacterium construction method. The sinonovacula constricta C1q gene, the encoded protein, the cloning method of the sinonovacula constricta C1q gene and the recombinant sinonovacula constricta C1q gene engineering bacterium construction method are characterized in that a sequence of the sinonovacula constricta C1q gene is as shown in SEQ ID NO.1; the cloning method of the sinonovacula constricta C1q gene comprises the steps of designing nested primers of 3'RACE according to an EST (expression sequence tag) sequence homologous with the C1q gene and amplifying a full gene length by adopting a 3'RACE technology; an amino acid sequence of the encoded protein of the sinonovacula constricta C1q gene is as shown in SEQ ID NO.2, and a sinonovacula constricta C1q protein is amplified by a primer respectively containing a BamH I site and an Xho I site; a target gene obtained by cloning is inserted into a vector to obtain a recombinant plasmid, and the recombinant plasmid is subjected to inducible expression and then is purified to obtain a gene engineering bacterium. The sinonovacula constricta C1q gene has the advantages of combining capacity for lipopolysaccharide, peptidoglycan and mannan and obvious agglutination effects for escherichia coli, vibrio anguillarum and vibrio parahaemolyticus.

The invention relates to the field of microbial disease control and in particular relates to a probiotic with bacteriostatic ability. The probiotic is Enterobacter ludwigii-PC88, is collected in China General Microbiological Culture Collection Center (CGMCC) on June 24, 2013, and has a collection number of CGMCC No.7814. The probiotic is separated from intestinal tracts of healthy flounders. The pure culture of the probiotic has relatively strong inhibitory effects on aquatic pathogens such as vibrio splendidus, vibrio harveyi, vibrio parahaemolyticus, pseudomonas aeruginosa and vibrio anguillarum, also can inhibit growth of known human pathogens such as tritirachium album, staphylococcus aureus and klebsiella pneumoniae, and has resistance to ampicillin. The pure culture or composition of the probiotic can be used for control of aquaculture diseases and development of novel antibacterial materials, and has good application prospects.

The invention relates to the field of vaccinology, and particularly relates to a vaccine utilizing vibrio anguillarum flagellin and an application thereof. According to the invention, vibrio anguillarum is taken as a template and F1/R1 is taken as a primer to obtain a PCR (polymerase chain reaction) amplification product which is connected with pET259, and after connection transformation, plasmid is split to be mixed with an adjuvant, so as to obtain the vaccine utilizing vibrio anguillarum flagellin, wherein the primer is F1: 5'-CCCGGGATGCCTAAGGAGATCAATATG-3' and R1: 5'-CCCGGGACCCAATAGACTAAGAGCT-3'. The vaccine has the immune protection efficiency reaching 78% on vibrio anguillarum.

The invention discloses an anti-Vibrio anguillarum yolk antibody, and the preparation method, the titer detection method and the application thereof. The invention is characterized in that the anti-Vibrio anguillarum yolk antibody is obtained by immunizing laying hens with Vibrio anguillarum as antigen, collecting highly-immunized eggs and performing separation and purification of yolk of the immunized eggs. The method comprises a step of preparing a Vibrio anguillarum antigen and inactivating the antigen, a step of performing basic immunization once in laying hens and secondary immunization for two to three times and collecting eggs, and a step of performing centrifugal separation, salting-out and dialysis to obtain the final product. The titer of the anti-Vibrio anguillarum yolk antibody is determined by indirect enzyme-linked immunosorbent assay. The anti-Vibrio anguillarum yolk antibody can be used for prevention and treatment of diseases caused by Vibrio anguillarum infection and can also be used for development of Vibrio anguillarum test kits and immunoreaction and diagnosis in immunology. The invention has the advantages that the yolk antibody is higher in purity and has a significant inhibitory effect on Vibrio anguillarum; the extraction of highly-immunized yolk is cost-efficient and convenient, and is adapted to low-cost and mass production; and the preparation method is simple and feasible.

The invention relates to the field of microbial disease control and in particular relates to a probiotic separated from intestinal tracts of turbots and an application thereof. The probiotic is Leclercia adecarboxylata P115, is collected in China General Microbiological Culture Collection Center (CGMCC) on June 24, 2013, and has a collection number of CGMCCNo.7813. The probiotic is separated from the intestinal tracts of healthy turbots. The probiotic and the pure culture thereof have obvious bacteriostatic effects on tritirachium album, klebsiella pneumoniae, staphylococcus aureus, vibrio parahaemolyticus, vibrio harveyi and vibrio anguillarum. The pure culture or composition of the probiotic can be used for control of aquaculture diseases and development of novel antibacterial materials, and has good application prospects.

The invention discloses a method for preparing a modified carbon nanotube-chitosan composite material. The method comprises the following steps: (1) modifying multi-walled carbon nanotubes with dopamine, so as to obtain dopamine modified carbon nanotubes; (2) subjecting a chitosan solution and a glutaraldehyde solution to a stirred and mixed reaction, and carrying out dialysis in distilled water, so as to obtain a chitosan-glutaraldehyde solution; preparing a 2mmol/L modified carbon nanotube solution from the modified carbon nanotube material obtained in the step (1), adding the modified carbon nanotube solution into the chitosan-glutaraldehyde solution, carrying out a reaction, then, adding a sodium borohydride solution, carrying out dialysis in distilled water, and then, carrying out standing, suction filtration, washing and drying. According to the method, the operation is simple, and the prepared modified carbon nanotube-chitosan composite material is high in chitosan graft amount, which is 71.78%, and presents excellent bacteriostatic performance to Escherichia coli, Staphylococcus aureus and Vibrio anguillarum, thereby having broad-spectrum bacteriostatic performance.

The invention relates to an aquaculture animal disease prevention and control technology, specifically to a Vibrio anguillarum 01 serotype inactivated vaccine, a preparation method and a use method thereof, wherein the vaccine is Vibrio anguillarum 01 serotype, the Vibrio anguillarum 01 serotype strain is preserved in China General Microbiological Culture Collection Center (CGMCC) on December 7, 2012, and a preservation number is CGMCC No.6942. With the inactivated vaccine, breeding fishes can be effectively stimulated to produce specific immune response, immunity of breeding animals can be effectively enhanced, and susceptible fishes can be protected from pathogenic Vibrio anguillarum infection. In addition, the Vibrio anguillarum 01 serotype inactivated vaccine preparation method has characteristics of low raw material cost and safety on animals and environment.

The invention relates to a preparation method of a microbial bactericide, which belongs to the field of microbial fermentation industry. The invention discloses a method for preparing Vibrio anguillaris bactericide by using Pseudoalteromonas sp., which consists of (1) slant activation and culture of strains, (2) liquid fermentation culture, (3) crude fermented liquid Mention, (4) steps such as the preparation of bacteriostatic agent are formed. The method of the invention is simple, the culture medium is cheap and easy to prepare, and the liquid culture technology is adopted, which is beneficial to enlargement and industrialized production, and has stable properties and good application value.

The invention relates to the technical field related to elimination of thalli, and particularly provides a preparation for strongly eliminating vibrios and a preparing method of the preparation. On one hand, provided is the preparation for strongly removing vibrios, wherein the preparation comprises the components in percentage by weight: 65-85% of a first guanidine derivative, 2-12% of a second guanidine derivative and the balance being ethanol, wherein the first guanidine derivative is polyhexamethylene guanidine salt and/or polyhexamethylene biguanide salt; and the relative molecular weightof the second guanidine derivative is 500-1500. The vibrios can be effectively removed by utilizing polyhexamethylene guanidine hydrochloride and chlorhexidine hydrochloride under the specific content. The preparation provided by the invention has a strong removal effect on pathogenic vibrios, such as vibrio parahaemolyticus, vibrio anguillarum, vibrio alginolyticus, vibrio cholerae, vibrio fluorescens and the like, has a good cracking effect on aeromonas hydrophila, aeromonas punctata, escherichia coli, salmonella and other gram-negative bacteria, and has a quite high removal rate after being used for 48 hours.