132 results about "Pathogenic vibrio" patented technology

The invention discloses an application of bdellovibrios in removing pathogenic vibrio in marine food products and the culture water. The bdellovibrio concentrated solution is added into the marine food products and/or the culture water so as to lead the concentration of the bdellovibrio to reach at least 10<2>pfu /ml. When the invention is applied before eating the marine food products, in the transportation process and in the culture water, the concentration range of bdellovibrio is respectively 10<4>-10<12> pfu /ml, 10<3>-10<11> pfu /ml, and 10<2>-10<6 > pfu /ml. More than 90% pathogenic vibrio carried by marine food products before eating and/or in the transportation process is cleared by adopting a biological method, and the pathogenic vibrio in the culture water can also be controlled within10cfu/ml. The invention is suitable for the pretreatment process before being eaten or processed, the transportation process and the culture process of marine food products, especially facilitating the reduction or elimination of chemical medicine residues such as antibiotics, thus fundamentally avoiding the occurrence of food poisoning of the marine food products.

The invention discloses a microbial preparation for improving an aquatic product culture environment and a preparation method thereof. A strain with optimal nitrite degradation effects is screened and separated from a mud sample in Tongan and Haicang ports of Xiamen city of the Fujian province, another strain with optimal nitrite degradation effects is screened and separated from a mud sample in the intertidal zone of Ningde city of the Fujian province, and the two strains respectively are Rhodopseudomonas palustris PSB20 and Bacillus subtilis ND04. The microbial preparation prepared from the two strains can degrade ammonia nitrogen and nitrous acid in an aquatic product culture environment, has good decomposition effects on residual bait organic matters, has a certain effect of inhibiting pathogenic vibrio bacterium growth in culture water, and has a good production application prospect. The microbial preparation has the characteristic of simple preparation processes.

The invention provides bacillus subtilis (Bacillussubtilis) shou003, an anti-vibrio protein and a preparation method and applications of the bacillus subtilis shou003 and the anti-vibrio protein. The preparation method of the anti-vibrio protein comprises the following steps: screening out bacillus subtilis shou003 (preserved in China Center for Type Culture Collection with a number of CCTCC No.: M2013571 on November 13, 2013) from intestinal tract of a healthy large yellow croaker; and then extracting the anti-vibrio protein from the fermentation broth of bacillus subtilis shou003, wherein the amino acid sequence of the anti-vibrio protein is shown as SEQ ID No.: 1. The bacillus subtilis shou003 and the anti-vibrio protein show good effects on inhibiting aquatic pathogenic bacteria, in particular pathogenic vibrio; in addition, the bacillus subtilis shou003 shows outstanding tolerance to temperature, NaCl, gastric juice, intestinal juice and cholate and can be widely applied to the prevention of germs during aquaculture; on that basis, the optimal fermentation culture method of the bacillus subtilis shou003, and a preparation method of the anti-vibrio protein are provided.

The invention provides a kit capable of detecting 10 pathogenic vibrios simultaneously based on a fluorescence probe melting curve method and a method capable of detecting the 10 pathogenic vibrios simultaneously. The kit comprises a universal primer, a fluorescence probe and hybridization connecting probes designed according to specific genes of the 10 pathogenic vibrios. The method capable of detecting the 10 pathogenic vibrios simultaneously includes the steps that the DNA of dubious bacteria extracted by a sample to be detected is used as a template, and hybridization connection reacting, fluorescence PCR amplification treatment and melting curve analysis treatment are carried out in sequence through the kit by means of the fluorescence probe melting curve method. According to the kit and the detection method, based on the multiple-connecting probe amplification technology, the 10 pathogenic vibrios can be detected simultaneously through the real-time fluorescence PCR melting curve method, hence, the detection period of the pathogenic vibrios is effectively shortened, and the detection efficiency of the pathogenic vibrios is improved.

The invention discloses a quadruple fluorescent PCR kit for detection of pathogenic vibrios in a water body and a detection method. According to the method, innovative bacterium enrichment reagent technology, DNA extraction reagent technology, specific fluorescent probe hybridization PCR detection technology and multiplex PCR technology are employed, bacterium enrichment culture of a sample is not needed, false positive interference is minimized as much as possible on the premise that high sensitivity is maintained, and four kinds of vibrios can be detected at one time. The invention mainly has the following beneficial effects: no need for bacterium enrichment of the sample in advance; high sensitivity and good specificity in detection of bacteria; reduction of the false positive incidence in conventional PCR amplification; and realization of rapid, accurate and specific detection of vibrio vulnificus, vibrio alginolyticus, vibrio mimicus and vibrio flurialis at one time.

The invention relates to a gene chip for detecting nine pathogenicity vibrios in marine products and a kit thereof. The gene chip comprises a solid phase carrier and an oligonucleotide probe, wherein the oligonucleotide probe includes one or more selected from the following nucleotide sequences: 1) DNA sequences selected from genes of a vibrio hollisae, a vibrio vulnificus, a vibrio cholera, a vibrio parahemolyticus, a vibrio harveyi, a vibrio alginolyticus, a vibrio furnissi, a vibrio mimicus and a vibrio damsel, 2) complementary DNA sequences of the DNA sequences selected in the DNA sequences in 1), 3) complementary RNA sequences of the selected DNA sequences in 1) or 2). The kit comprises the gene chips. The gene chip for detecting nine pathogenicity vibrios in marine products and the kit thereof provided by the invention are used for detecting the nine pathogenicity vibrios in marine products, and have the advantages of simplicity in operation, good sensitivity and strong repeatability.

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 provides a murine hybridoma cell line, a monoclonal antibody resistant to thermostable direct hemolysin of vibrio parahaemolyticus and secreted by the hybridoma cell line and a preparation method of the monoclonal antibody. The monoclonal antibody resistant to the thermostable direct hemolysin of the vibrio parahaemolyticus for mice is stably secreted by one hybridoma cell line T6D4 obtained by means of B lymphocyte hybridoma technique after TDH (thermostable direct hemolysin) of the vibrio parahaemolyticus is extracted and purified and the BalB/c mice are immunized, and is used for rapid detection of pathogenic vibrio parahaemolyticus. The antibody is IgG2a secreted by one stable murine hybridoma cell line and is capable of generating idiosyncratic reaction to TDH.

The invention discloses an ssDNA nucleic acid aptamer and application thereof to detect pathogenic vibrio alginolyticus. The nucleotide sequence of the ssDNA nucleic acid aptamer is 5'-GACGCTTACTCAGGTGTGACTCGCGTTTTATTGGTGTGGGGCTGGGGCGGTGGGTGGCTCTACTGGTTCCGTTCGAAGGACGCAGATGAAGTCTC-3'(SEQ ID NO:1). The ssDNA nucleic acid aptamer is specific for and highly sensitive to the trachinotus ovatus source pathogenic vibrio alginolyticus, and has no immunogenicity. The ssDNA nucleic acid aptamer is stable in structure and easy to modify, facilitates synthesis and preservation, and can rapidly and accurately detect and diagnose the trachinotus ovatus source pathogenic vibrio alginolyticus.

The invention belongs to the technical field of aquatic organisms and in particular relates to a quick detection method for scallop pathogenic vibrio splendidus. Total DNA (deoxyribonucleic acid) of bacterial genomes in a filtered seawater sample to be detected is taken as a template, fluorescent quantitative PCR (polymerase chain reaction) amplification is carried out by a bacterial 16SrDNA conserved region and a specific primer of a pathogenic vibrio splendidus metalloproteinases gene, and the scallop pathogenic vibrio splendidus in the culture environment is quantitatively analyzed and detected; the detection method comprises the steps of extracting the DNA of the bacterial genomes and carrying out fluorescent quantitative PCR detection. The method has the advantages that the quickness is achieved, the specificity is good, the bacterial density detection range is wide, and a quantitative result can be close to a real situation.

The invention discloses a general nucleic acid isothermal amplification detection kit for pathogenic vibrios of mariculture animals. The detection kit comprises a grinding liquid tube into which grinding liquid is filled, a nucleic acid extracting solution tube A into which solution of sodium acetate is filled, a nucleic acid extracting solution tube B into which absolute ethanol is filled, a nucleic acid extracting solution tube C into which 70 mass percent ethanol solution is filled, a tris-hydrogen chloride ethylene diamine tetraacetic acid (TE) buffer solution tube into which TE buffer solution is filled, a uracil-DNA-glycosylase (UNG) tube into which uracil-DNA-glycosylase is filled, a loop-mediated isothermal amplification (LAMP) reaction liquid tube into which LAMP reaction liquid is filled, a bacillus stearothermophilus deoxyribonucleic acid (Bst DNA) polymerase tube into which Bst DNA polymerase is filled, a color-developing agent tube into which a nucleic acid dye SYBR Green I is filled, a positive control nucleic acid tube into which positive DNA of the vibrios is filled and a negative control tube into which sterilized double distilled water is filled. The invention also discloses a method for detecting the pathogenic vibrios of the mariculture animals by utilizing the detection kit.

The invention relates to an immunomagnetic bead containing an OmpU antibody, preparation of the immunomagnetic bead and a method for capturing pathogenic vibrios and detecting the pathogenic vibrios through multiplex PCR by utilizing the immunomagnetic bead. PCR amplification of an OmpU gene is conducted by taking genomes of the vibrios as templates, pure OmpU recombinant protein is obtained to serve as an antigen to prepare the antibody, and the immunomagnetic bead containing the OmpU antibody is prepared through the coupling-washing process; the immunomagnetic bead can capture multiple vibrios, and then the vibrios are identified by multiplex PCR specific primers which are designed on the basis of a danJ gene. The immuomagnetic bead can simultaneously detect the vibrio parahaemolyticus, the vibrio cholera, the vibrio alginolyticus, the vibrio vulnificus and the vibrio mimicus at most and can be widely applied to environment and import and export food inspection. The immuomagnetic bead combines the specificity of immune separation and the high efficiency of PCR amplification, has the capacities of separation, concentration and enrichment, can efficiently separate and enrich the low-concentration pathogenic bacteria in a sample, reduces or eliminates inhibition and influences of the sample to PCR, improves detection limitation and has the very good application prospect.

The invention relates to a method for preparing a bdellovibrio preparation by an enrichment culture method, belongs to the field of biotechnology and relates to an enrichment culture method of variousbdellovibrio, host bacteria culture and production methods of biological preparations thereof. The method is characterized in that a bdellovibrio system naturally formed in aquaculture water culturedby the enrichment culture method is used as a source of the bdellovibrio of the biological preparation, and pathogenic vibrio in the aquaculture water is purified, cultured, centrifuged, inactivatedand then used as a source of the host bacteria for the production of the bdellovibrio preparation to produce the biological preparation containing various bdellovibrio. The bdellovibrio preparation produced by the method has the advantages of high activity, rapid action and broad cleavage of pathogenic bacteria in the water due to the various bdellovibrio.

The invention relates to a gene chip capable of simultaneously detecting various vibrios and a method for detecting the vibrios, and provides species-specific genes of four common pathogenic vibrios in the aquatic environment to encode an oligonucleotide probe sequence of an hsp60 gene and an oligonucleotide probe sequence of a toxR gene, wherein the four common pathogenic vibrios include vibrio alginolyticus, vibrio parahaemolyticus, vibrio vulnificus and vibrio haeveyi. On the basis of the base complementary and pairing principle, by combining the method with an optimized ordinary PCR and fluorescence labeled PCR reaction system, the aim of detection is achieved through oligonucleotide probes fixed to a solid-phase aldehyde chip. The invention provides the detecting method which is high in flux, specificity and sensitivity, rapid and efficient for the bottleneck of a current method for detecting pathogenic microorganisms.

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 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.

The invention relates to a powdery preparation of bacillus pumilus and bdellovibrio and belongs to the technical field of feed additives. The powdery preparation contains the bacillus pumilus and the bdellovibrio of which the preservation number is CCTCC M 2013240, wherein the bdellovibrio is one or more of bdellovibrio bacteriovorus, bdellovibrio stolpii and bdellovibrio starrii. The invention also provides application of the powdery preparation in preparation of a feed additive for treating a disease caused by vibrio. The powdery preparation has the advantages of effectively treating the disease caused by the vibrio in a broad-spectrum manner, inhibiting the growth of pathogenic vibrio in the intestinal tract of a prawn and reducing occurrence of a vibrio disease of the prawn when being added into a feed for the prawn, also promoting the growth of the prawn, lowering the cultivation cost and being worthy of wide application in aquaculture.

The invention provides vibrio parahaemolyticus bacteriophage lyase PCNP-lys, and the amino acid sequence of the PCNP-lys is shown in SEQ ID NO:1. The invention further provides an antibacterial agentwhich comprises main active components of the vibrio parahaemolyticus bacteriophage lyase PCNP-lys, a carrier with a PCNP-lys expression element, an expression cassette with the PCNP-lys expression element or at least one of host cells with the PCNP-lys expression element. The invention further provides a preparation method of the vibrio parahaemolyticus bacteriophage lyase PCNP-lys. The vibrio parahaemolyticus bacteriophage lyase PCNP-lys provided by the invention is capable of splitting multiple pathogenic vibrios, and makes a basis for substitution therapies of drug-resistant vibrio parahemolyticus.