41 results about "Vibrio splendidus" patented technology

The invention relates to the technical field of functional microbiological screening and provides novel Bacillus licheniformis DN29 which is collected under CCTCC NO: M2015482. The Bacillus licheniformis DN29 obtained by screening can effectively inhibit pathogenic bacteria such as Vibrio splendidus and pseudoalteromonas, can reduce disease probability of bred animals and can serve as a feed additive to remarkably increase feed utilization rate of the bred animals and promote growing of the animals. The Bacillus licheniformis DN29 can serve as a probiotic to be applied in the process of rearing and producing Stichopus japonicus, has remarkable promoting effect on growing of Stichopus japonicus and can effectively improve immunity of and resistance, to Vibrio splendidus, of Stichopus japonicus.

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 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 a gold-labelled nucleic acid test strip for quickly detecting aquatic diseases. The gold-labelled nucleic acid test strip comprises a sample pad, a gold-labelled pad containing probe Probe1-AuNPs composite, a nitrocellulose membrane and a water absorbent pad; a detection line and a quality control line which are separated from each other are respectively adsorbed on the nitrocellulose membrane; the detection line is composed of a probe2, and the quality control line is composed of the probe 1'; the probe1 and the probe2 are respectively designed by selecting a characteristic conserved segment from a vibrio splendidus toxicity gene sequence and using the 5' end and 3' end of the characteristic conserved segment, and then a complementary sequence Probe1' is designedaccording to the probe1. A nano-gold nucleic acid probe is used as the label in the development of the gold-labelled nucleic acid test strip, and the vibrio splendidus is detected through a thin-filmchromatography measure, thereby providing simple, convenient and efficient detection method for the disease control of the vibrio splendidus.

The invention discloses a vibrio splendidus colloidal gold immunochromatograohic assay test strip capable of rapidly detecting pathogenic bacteria of stichopus japonicus 'bark rot' and a preparation method thereof. The test strip comprises a bottom plate, wherein a sample pad, a gold-labeled pad, a nitrocellulose membrane and a water absorbent pad are sequentially adhered onto the bottom plate from one end from top to bottom in a stepped manner; a colloidal gold labeled vibrio splendidus flagellin FlgT monoclonal antibody is sprayed onto the gold-labeled pad; a detection line a coated with a vibrio splendidus polyclonal antibody and a quality control line b composed of goat anti-rat IgG are arranged on the nitrocellulose membrane; and the vibrio splendidus flagellin FlgT monoclonal antibody is prepared by immunizing mice by hybridoma cells 4A5 with a collection number of CGMCC No. 14311, and purifying mouse ascites obtained.

The invention provides a strain of Bacillus subtilis ZF003 (Bacillus subtilis s), which is preserved in China General Microbiological Culture Collection Center and is assigned the accession number ofCGMCC No.15645, preservation date being Apr 26th, 2018. The invention also discloses an application of the Bacillus subtilis, wherein the Bacillus subtilis is applied to cultivation of stichopus japonicas, comprising two processes: (1) fermenting a stichopus japonicas feed with the Bacillus subtilis; (2) spreading a microbial liquid of the Bacillus subtilis in a breeding pool of the stichopus japonicas. The strain, on one hand, can secrete the lipase extracellularly, so that fat in the feed is decomposed by the lipase; and on the other hand, the strain can inhibit the main pathogenic bacteria,Vibrio splendidus, of skin ulcer syndrome of the stichopus japonicas.

The invention discloses a feed additive for stichopus japonicus, a preparation method and a compound feed containing the feed additive. The feed additive for the stichopus japonicus comprises the following raw materials: rhubarb, baical skullcap root, Chinese gall, honeysuckle flower, wild chrysanthemum, indigowoad root and a synergistic agent. The preparation method comprises the following stepsof: weighing the raw materials in a weight ratio; crushing all the raw materials; and putting all the crushed raw materials into a mixer sequentially, and mixing uniformly. The compound feed containing the additive comprises 1 to 3 weight percent of additive. The technical scheme provided by the embodiment of the invention has the advantages that: the additive is prepared from Chinese medicinal herbs, does not have medicinal tolerance and toxic or side effects, can improve the growth speed of the stichopus japonicus obviously, improve the immunological activity of the stichopus japonicus and enhance the disease-resistant capacity of the stichopus japonicus, and has extremely high resistance to vibrio splendidus serving as main pathogenic bacteria. The stichopus japonicus is difficult to sicken, the survival rate is improved obviously, and the production benefit is improved obviously.

The invention discloses a method for quickly detecting pathogen of skin ulcerative syndrome in Apostichopus on site and a kit thereof. In the method, two pairs of primers, particular operation steps and the kit aiming at the vibrio splendidus are designed by using a loop-mediated isothermal amplification (LAMP) method. The DNA sequences of the reaction primer are as follows: VS-F3 is 5'-TGGTCCCCCCATATTCAGAC; VS-B3 is 5'-AAGTACCCCGAGGAAGAGAA; VS-FIP is 5'-GCCCCGTAACCGACACATCATTTTAGGATATCACGTGTCCCGC; and VS-BIP is 5'-GGCACTTTCCAGAGCCTTCGTTTTTAGATTCCGAAAGTAGCGGC. By using the method, expensive instrument and equipment or long detecting time is not required, the pathogen can be detected even less pathogen exists. The method has the advantages of low cost, accurate detection and the like, is convenient to operate, real-time monitor can be performed, and the epidemic situation can be discovered in time, the incidence trend of the epidemic disease is reduced, the pathogen can be prevented from distributing in early stage, and more loss brought to raisers is avoided.

The invention provides a gene chip for detecting various seawater cultured animal pathogenic bacteria and its application. The pathogenic bacteria include: Vibrio anguillarum, Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio harveyi, Vibrio fischeri, Vibrio splendidus, Vibrio riverina, Vibrio vulnificus, Vibrio cholerae non-O1, Vibrio mimicus, Vibrio salmonicida, Aeromonas hydrophila, Aeromonas sobria, Aeromonas salmonicida, Edward tarda Bacillus, Edwardsiella catfish, Renalobacter salmonidum, Mycobacterium marinum, Pseudomonas, Streptococcus.

The invention relates to a microecological preparation used for preventing and treating apostichopus japonicus putrid skin syndrome and a preparation method of the microecological preparation and belongs to the field of aquatic microorganisms. Active ingredients of the microecological preparation are thalli of bacillus megatherium H1 and extracellular metabolites of the bacillus megatherium H1; the bacillus megatherium strain is separated from intestinal tract excrement of apostichopus japonicus which is healthily cultivated in Penglai, Shandong, detection of safety, bacteriostatic activity and toxicity and artificial antagonistic vibrio splendidus experiments verify that the bacillus megatherium H1 is antagonistic to vibrio splendidus, young apostichopus japonicus is effectively protected, the bacillus megatherium H1 can be used for preventing and treating apostichopus japonicus 'putrid skin syndrome' caused by vibrio splendidus and survival rate of an apostichopus japonicus seedling protecting stage is increased.

The invention provides a combined inactivated vaccine of photobacterium damselae and vibrio splendidus of pampus argenteus. The preservation number of the photobacterium damselae is CGMCC NO.16907. The combined inactivated vaccine is simple in preparation process, stable in yield and low in cost, and a large amount of inactivated vaccines with a high immune effect can be obtained. The immune protection rate of the inactivated vaccine prepared by the method reaches 92.5%; the vaccine is a pure biological preparation, and is safe and environmentally friendly, drug residues and environmental pollution are avoided, the occurrence and prevalence of the photobacterium damselae and vibrio splendidus diseases of the pampus argenteus can be fundamentally restrained, and technical support is provided for large-scale preparation of the bigeminy inactivated vaccine for the photobacterium damselae and vibrio splendidus diseases of the pampus argenteus.

The invention discloses a gene chip for detecting pathogenic vibrio flora in sea cucumber, shrimp and shellfish culture regions, and a use method. The gene chip comprises a chip carrier, wherein chemically modified nucleotide probes for detecting vibrio parahaemolyticus, vibrio alginolyticus, vibrio anguillarum, vibrio harveyi, vibrio vulnificus and vibrio splendidus are immobilized on the chip carrier. The use method comprises the following steps: DNA extraction of a sample to be detected, PCR amplification of the sample, hybridization of a PCR product with a chip, processing after hybridization of the chip, scanning of hybridization chip, and interpretation of results. The gene chip significantly improves the detection specificity by specific probes designed for each vibrio variation region sequence, improves the capability of identifying pathogenic vibrio flora of environments and organisms of the sea cucumber, shrimp and shellfish culture regions, and also greatly shortens the detection time.

The invention discloses sinonovacula-constricta I-type lysozyme-2 genes, encoded protein and an construction method and application of recombinant sinonovacula-constricta I-type lysozyme-2 gene engineering bacteria. The sinonovacula-constricta I-type lysozyme-2 genes are characterized in that sequences of the sinonovacula-constricta I-type lysozyme-2 genes are shown in SEQID NO.1; a clone method of the sinonovacula-constricta I-type lysozyme-2 genes is characterized in that nested primers of 3'RACE are designed according to expression sequence tag (EST) sequences homologous with the sinonovacula-constricta I-type lysozyme-2 genes, and the full length of the genes is amplified with the 3'RACE technology; amino acid sequences of the sinonovacula-constricta I-type lysozyme-2 gene encoded protein are shown in SEQID NO.2, and the sinonovacula-constricta I-type lysozyme-2 protein is amplified with primers containing BamH I loci and Xho I loci respectively; target genes obtained after cloningare inserted into carriers to obtain recombinant plasmids, the recombinant plasmids are subjected to inducible expression and then are purified, and the gene engineering bacteria are obtained. The gene engineering bacteria have the inhibiting effect on vibrio harveyi, vibrio parahaemolyticus and vibrio splendidus.

The invention discloses a genetic molecular marking combination for anti-disease stichopus japonicas breeding and application of the genetic molecular marking combination and belongs to the field of aquatic product genetic breeding. The genetic molecular marking combination consists of three SNP (Single Nucleotide Polymorphism) markers SNP88, SNP112 and SNP126 related to anti-vibrio splendidus infection of stichopus japonicas, and the three marked dominant genotypes are respectively SNP88 (CC), SNP112 (AA) and SNP126 (TT). A breeding method with the genetic markers comprises the following operation steps: (1) extracting parent DNA (Deoxyribonucleic Acid) and typing; (2) selecting parents; (3) performing filial generation seedling breeding. By adopting the genetic molecular marking combination, effective molecular markers for improving the economic property of anti-vibrio splendidus infection of stichopus japonicas are provided, the genetic markers are adopted for auxiliary marker selection of stichopus japonicas, anti-disease breeding can be carried out, the anti-vibrio splendidus infection of the stichopus japonicas can be improved, the morbidity of skin ulcer syndromes of the stichopus japonicas can be substantially reduced, and the combination has great significances in increasing the breeding survival rate and the breeding benefits.

The invention discloses a hybridoma cell strain 4A5 and a vibrio splendidus flagellin FlgT monoclonal antibody. The hybridoma cell strain 4A5 is collected in China General Microbiological Culture Collection Center with the collection number of CGMCC No.14311. The hybridoma cell strain 4A5 is obtained by utilizing recombinant protein FlgT as an antigen to immunize a mouse, taking spleen cells of the mouse to be fused with SP2/0 cells, adopting a limiting dilution method to obtain the monoclonal antibody, and adopting an ELISA (Enzyme Linked Immunosorbent Assay) method to screen positive hybridoma cells; and the vibrio splendidus flagellin FlgT monoclonal antibody secreted by the hybridoma cell strain 4A5 has more specificity, and can be applied in the field of immunodetection.

The invention provides a bacillus thuringiensis strain XW008. The strain is collected in the China General Microbiological Culture Collection Center, has a collection number of CGMCC No. 15647, and has a collection date of April 26, 2018. The invention further discloses application of the bacillus thuringiensis. The bacillus thuringiensis is applied to culture of stichopus japonicas, and the application comprises two methods: (1) fermenting stichopus japonicas feed by adopting the bacillus thuringiensis (2) splashing bacterium liquid of the bacillus thuringiensis into a stichopus japonicas culture pond. On one hand, the strain is capable of secreting extracellular cellulase, and the cellulase is capable of decomposing cellulose in the feed; on the other hand, the main pathogen, namely vibrio splendidus, of skin ulcerative syndrome of stichopus japonicas (also called rotten-skin disease) can be effectively inhibited.

The invention discloses a primer and a method for rapid isothermal amplification detection of vibrio splendidus based on a microfluidic chip and belongs to the technical field of microbial detection. According to the method, a loop-mediated isothermal amplification (LAMP) system containing three pairs of primers and a reaction pool array microfluidic chip detection method are adopted, three pairs of primers designed for a specific region in a vibrio splendidus arsenic-resistant gene arsB are finally determined through LAMP primer design and screening, and specific sequences are shown as SEQ ID NO.1/SEQ ID NO.2; SEQ ID NO.3/SEQ ID NO.4; SEQ ID NO.5/SEQ ID NO.6. The method is used for isothermal amplification detection of the vibrio splendidus based on the microfluidic chip, has high amplification efficiency and specificity, is simple and rapid in operation process, makes results stable and reliable, can be well used for early rapid detection and timely prevention and control of the vibrio splendidus and has a good application prospect.