201 results about "Vibrio species" patented technology

The invention discloses a detection kit and a detection method for 9 species of pathogenic organisms in marine products. The kit comprises Taq DNA polymerase with a concentration of 5U / muL and a PCR reaction solution, wherein the PCR reaction solution contains 10 millimols of Tris.HCl, 50 millimols of KCl, 25 millimols of MgCl2, 2.5 millimols of dNTP and primer pairs of the 9 species of pathogenic organisms. The kit and the method can synchronously detect the 9 species of pathogenic organisms and can detect salmonella with a concentration of 11 CFU / mL, staphylococcus aurei with a concentration of 2,000 CFU / mL, monocytogenes with a concentration of 120 CFU / mL, enterohemorrhagic Escherichia coli O157:H76 with a concentration of CFU / mL, comma bacillus with a concentration of 220 CFU / mL, Vibrio parahaemolyticus with a concentration of 60 CFU / mL, Vibrio vulnficus with a concentration of 9 CFU / mL and Vibrio alginolyicus with a concentration of 230 CFU / mL. The detection method short in detection time and simple and quick in operation, and can save a large amount of labor and financial resources and meet requirements for quick detection.

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 invention discloses an oligonucleotide primer for detecting common pathogenic bacteria by adopting a fluorescent quantitation PCR (Rich Client Platform) technology, a method thereof for detecting common pathogenic bacteria and the application thereof. The method comprises the following steps of: providing 10 pairs of specific oligonucleotide primer sequences at annealing temperature of 50-60 DEG C without differing 5 DEG C; and simultaneously, quickly, accurately and effectively identifying and quantificationally detecting various pathogenic bacteria at the same time. A detection range comprises bacillus cereus, enterobacter sakazakii, vibrio parahaemolyticus, enterohemorrhagic escherichia coli O157, salmonella, Listeria monocytogenes, Shigella, campylobacter jejuni, pseudomonas aeruginosa, klebsiella pneumoniae, and the like. The invention also can be used for the fields of disease diagnosis, environmental monitoring, water-quality and food supervision and detection, food poisoning pathogenicbacteria detection, bacteriological classification, epidemiological investigation, biological agent detection, and the like, is convenient, quick, accurate and effective and has wide application range.

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.

Disclosed is a method for detecting the species of a bacterium in a simple manner. Specifically disclosed is a method for detecting the species of a bacterium, which can detect at least one bacterial species selected from Staphylococcus species, Streptococcus species, Klebsiella species, Escherichia species, Mycobacterium species, Legionella species, Vibrio species, Bacillus species, Neisseria species, Campylobacter species, Chlamydia species, Chlamydophila species, Mycoplasma species, Listeria species, Salmonella species and Yersinia species, and which comprises the steps of: a) contacting a sample suspected to contain a nucleic acid derived from the bacterium species to be detected with a nucleic acid molecule capable of hybridizing with at least a part of a DnaJ gene derived from the bacterial species; and b) detecting the presence or absence of the hybridization between the nucleic acid molecule and the nucleic acid contained in the sample.

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 discloses an agarase and an acquisition method of an encoding gene thereof. The acquisition method utilizes various PCR methods to acquire the agarase gene aga41 from marine bacteria vibrio natriegens (the preservation number is CGMCC No.2428) for the first time and breaks through the conventional method which screens the agarase gene by constructing a library. The agarase gene aga41 belongs to a GH50 family in a beta-agarase. The whole length of the agarase gene aga41 is 2,973bp. 990 amino acids are encoded on the agarase gene aga41. The highest sequence similarity of the agarase gene aga41 with the existing agarase is 49 percent. The agarase gene aga41 is a novel beta-agarase. An activated recombinant agarase which is obtained by cloning the agarase gene of the invention on an expression vector pET28b, constructing a recombinant escherichia coli Rosetta bacterial strain and carrying out heterologous expression can be applied to the industrial production.

The invention relates to a complex enrichment medium used for salmonella, vibrio parahaemolyticus and comma bacillus and a method for preparing the same. The formulation components of the complex enrichment medium in weight portion are as follows: 0.5 to 1.5 portions of buffer peptone water, 1000 portions of distilled water, 1.0 to 5.0 portions of glucose, 1.0 to 5.0 portions of mannitol, 0.03 to 0.06 portion sodium pyruvate, 0.5 to 2.5 portions of anhydrous sodium sulfite, 1.0 to 10.0 portions of sodium citrate, 5.0 to 25.0 portions of sodium chloride, 1 to 5 portions of cholate, and 0.0005 to 0.0025 portion of potassium tellurite. The method comprises the following steps: firstly, adding the buffer peptone water and the like into the distilled water, and sterilizing the mixture after heating and melting; and secondly, cooling the mixture to 50 DEG C, and then adding the potassium tellurite into the mixture to be mixed evenly. The complex enrichment medium can integrate two steps of primary enrichment and selective enrichment of bacterial detection to shorten the enrichment time to 24 hours, can perform enrichment on three target pathogens at the same time, and can inhibit the growth of other microorganisms.

The invention relates to an aquaculture composite microbial agent, which comprises the following agents by weight: 5-10 parts of Bacillus subtilis, 0.5-4 parts of Bacillus licheniformis, 1-6 parts of bacillus megaterium, 0.2-1 part of Pseudomonas gelatinasa, 0.5-3 parts of Rhodopseudomonas capsulata, and 2-8 parts of Bdellovibrio bacteriovorus. The invention also relates to application of the aquaculture composite microbial agent in improvement of water quality and / or control of aquatic organism diseases.

The invention relates to a method and treatment agent for preventing and controlling blue-green algae. A cyanophage bacterium S agent is a culture solution with the solid content of 0.5-2 percent, which is prepared from spirulina powder, NaCl and purified water; a biological substrate modified agent of K agent is prepared by compounding saccharomycetes, bacillus subtilis and cyanophage bdellovibro; and an algal removing water purifying agent of L agent is prepared from 40wt% of sodium carbonate, 50wt% of fourth generation of polyamidoamine and 10 wt% of sodium bentonite. The invention overcomes the defects of two methods by using beneficial bacterium which scrambles nutrition with the blue-green algae and cyanophage capable of phagocytosing the blue-green algae. The invention is a system engineering for preventing and controlling blue-green algae, which organically combines a biological method and a chemical method and can prevent and control blue-green algae comprehensively with proper cost and favorable practicability and stability; and the blue-green algae is controlled by using a microbial method at early stage with obvious implementation effect , since a cell wall of the blue-green algae is thinner and cyanophage can phagocytose algae quickly; in addition, since a water body has high eutrophication degree in summer, comprehensive treatment can achieve good effect.

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 present invention relates to a composition for preventing, improving, or treating aging, wherein the composition comprises a recombinant protein of flagellin, which is the constituent of Vibrio vulnificus flagella, fused with a pathogenic protein antigen, as an active component. According to the present invention, the recombinant protein of the present invention can improve external and internal aging-related malfunctions and enhance immunity. Also, the composition of the present invention can easily perform immunization through mucosal administration.

The present invention discloses a strain of marine Bacillus amyloliquefaciens B01, wherein the strain is preserved in the China general microbiological culture collection center on December 11, 2014, and has the preservation number of CGMCC No.10154. According to the present invention, the fermentation broth supernatant of the Bacillus amyloliquefaciens B01 can inhibit aquatic diseases caused by a variety of pathogenic bacteria such as Acinetobacter baumannii, Vibrio parahaemolyticus and the like, and the marine Bacillus amyloliquefaciens B01 has characteristics of no toxicity on human and animals, green environmental protection, aquatic pollution disease prevention and control effect, and good biological pesticide development prospects.

The invention relates to a gene chip for detecting important pathogenic bacteria in an aquatic product and a kit thereof. The gene chip comprises a solid phase carrier and an oligonucleotide probe; the oligonucleotide probe comprises one or more of the following nucleotide sequences: (1) DNA (Deoxyribonucleic Acid) sequences selected from proteus mirabilis 1, proteus vulgaris, salmonella, vibrio parahaemolyticus, vibrio cholerae, Listeria monocytogenes, staphylococcus aureus, streptococcus pyogenes, vibrio vulnificus, bacillus cereus cluster, Shigella and pathogenic Y. enterocolitica ail gene; (2) complementary DNA sequences of the DNA sequences selected in the (1); (3) complementary RNA (Ribonucleic Acid) sequences of the DNA sequences in the (1) or (2). The kit comprises the gene chip. The gene chip and the kit can be used for detecting the important pathogenic bacteria in an aquatic product, are convenient to operate, high in precision and strong in repeatability.

The invention provides a composite microecological feed additive capable of increasing the freshwater fish growth rate and application of the composite microecological feed additive. The additive comprises the following components: aspergillus niger solid fermentation compound enzyme, bacillus subtilis, bacillus cereus, cellulose, xylanase, neutral protease and phytase. The composite microecological feed additive disclosed by the invention is added with the solid fermentation compound enzyme, so that the utilization rate of the freshwater fish to baits is effectively increased, the growth of fishes is promoted, the breeding of pathogenic bacteria including aeromonas hydrophila, vibrio alginolyticus and the like in a water body can be inhibited due to the addition of fungicides, the occurrence possibility of diseases of saprolegniasis, white skin diseases, bacillary enteritis and the like is reduced; compared with similar additives, the additive disclosed by the invention has the advantages of being safe, high in efficiency, special, free of residues, free of side effects and the like; the composite microecological feed additive can be used for improving and keeping the good growth state of the freshwater fish and effectively increasing the freshwater fish growth rate.

The invention provides a method for preparing high-quality biological organic fertilizer through municipal sewage sludge. The method includes the steps of raw material preparing, fermented material preparing, sterilization, anaerobic fermentation, fermentation promoter injection, aerobic fermentation and finished product preparing. Anaerobic fermentation agents used in the anaerobic fermentation step contain methane thermophilic micrococcocci, lactobacillus planetarium, metallophilic thermoanaerobacter ethanolicus and white high-temperature actinomycetes. Aerobic fermentation agents used in the aerobic fermentation step contain trichoderma reese, lactobacillus delbruckii and vibrio proteolyticus. According to the prepared bio-organic fertilizer, the number of fecal coliform bacteria ranges from 23 to 26 per g, the death rate of ova of roundworm ranges from 99.2% to 99.8%, and the number of effective living bacteria ranges from 440 millions CPF / g to 52 millions CPF / g.

The present invention provides one kind of antibiotic peptide and its coding polynucleotides and use. The antibiotic peptide contains the amino acid sequence from site 1 to site 25 in the sequence table <400>2 of the sequence table. The polynucleotides coding the antibiotic peptide include the nucleotide sequence of amino acid sequence form site 1 to site 25, from site 1 to site 45 or from -22 site to site 45 in the sequence table <400>2 of the sequence table or their complementary sequences. The antibiotic peptide has obvious killing effect on several kinds of bacteria, and has lowest killing concentration smaller than 10 micro mol / L to algae dissolving vibrio, vibrio vulnificus, parahemolytic vibrio, Pasteuvella multocida, etc. and may be used in preparing antibiotic, especially that for preventing and treating bacterial diseases of aquatics.

The invention discloses a vibrio alginolyticus bacteriophage preparation and a preparation method and application thereof. Auxiliary agents such as vibrio alginolyticus bacteriophage Vp670 and trehalose are adopted for developing the corresponding bacteriophage preparation. After application, the bacteriophage preparation continuously exists in a water environment for a long time to keep quantitybalance of pathogenic host bacteria and bacteriophage, so that effect durability and low consumption are achieved as compared with those of antibiotics. The bacteriophage Vp670 preparation is free oftoxic and side effects, is a novel green biological sterilization preparation and can be used for prevention and control of vibrio alginolyticus infection of animals in marine aquaculture to promote healthy development of the aquaculture industry.

The invention relates to a V. alginolyticus outer-membrane protein W gene, process for preparing the same and uses. The invention provides a novel V.alginolyticus OmpW gene, and the use of V.alginolyticus and Vibrio vulnficus OmpW protein and coded sequence, which comprises, linking purified nucleic acid sequence encoding protein active polypeptides having V.alginolyticus OmpW to expression regulation sequence, forming V.alginolyticus OmpW protein expression carrier, transferring the expression carrier into host cells, forming the recombination cells of the V.alginolyticus Omp protein, culturing the recombination cells and separating.

The invention discloses a synthesis method of urea peroxide and applications of the urea peroxide in an antibacterial agent. The synthesis method comprises the following steps: adding hydrogen peroxide and urea into a reactor in proportion, and stirring at room temperature to dissolve hydrogen peroxide and urea; adding an additive into the reactor, and stirring at room temperature; and cooling andcrystallizing an solution in the reactor, filtering, and freeze-drying an obtained filter cake. The urea peroxide obtained by the invention is solid powder, is not easy to absorb moisture, is convenient to store and transport, is not easy to deteriorate, has a good sterilization effect, has good inhibition and killing effects on staphylococcus aureus, escherichia coli, bacillus sakazakii, salmonella typhimurium, listeria monocytogenes and vibrio parahaemolyticus, and is a potential broad-spectrum antibacterial agent.

The invention provides a preparation method of a mariculture fish multiple anti-idiotypic egg yolk antibody vaccine. The preparation method comprises the following steps of after inactivating mariculture fish pathogenic bacteria, immunizing mice and rabbits; after using six rabbit anti-pathogenic serums to be mixed with the inactivated mariculture fish pathogenic bacteria, immunizing laying hens;and screening the laying hens, collecting, refining and purifying an obtained coarse anti-idiotypic egg yolk antibody and the mariculture fish multiple anti-idiotypic egg yolk antibody vaccine, wherein the mariculture fish pathogenic bacteria are vibrio anguillarum, Edwardsiella tarda, vibrio alginolyticus, vibrio parahaemolyticus, vibrio vulnificus and streptococcus. The multiple anti-idiotypic egg yolk antibody vaccine provided by the invention can strengthen the resistance of marine fishes to various fish diseases, can prevent various marine fish diseases, can also perform immunization to the marine fishes through an oral way, is simple, convenient and feasible and is liable in obtaining materials at the aspect of preparing an anti-idiotypic egg yolk antibody, is convenient with aseptictreatment, is simple and convenient in preparation programs, low in cost and high in yield and accords with national policy orientation of energy conservation, environmental protection and sustainable development.

The present invention discloses a mutant blue fluorescent protein (BFP) exhibiting, at an aerobic or anaerobic system, larger fluorescent intensity than a BFPvv D7 of SEQ ID NO:2 derived from a wild type BFP, BfgV of SEQ ID NO:1, obtained from Vibrio vulnificus, wherein a set of mutation positions of the mutant BFP corresponding to SEQ ID NO:2 comprises position 176 or position 178. In a preferred embodiment, the set of mutation positions of the mutant BFP corresponding to SEQ ID NO:2 comprises a S176R mutation or a V178I mutation. Moreover, methods of using the blue fluorescent proteins from Vibrio vulnificus for fluorescence resonance energy transfer (FRET) and a blue fluorescent fish are also provided.

The present invention relates methods for the biosynthesis of tetrodotoxin (TTX) involving the steps of obtaining a culture possessing one or more of a Vibrio species, such as through a seed culture / inoculating the culture and tissue extract from a textrodotoxin-bearing organism in a fermenter medium, and isolating and purifying tetrodotoxin from said fermenter, resulting in a yield of TTX of about 0.5 g TTX / L in about 3 to 5 days, such TTX being at least 90% pure.

The invention is an ssDNA aptamer for specific recognition of vibrio vulnificus. The nucleotide sequence of the aptamer is a nucleotide sequence of a DNA fragment shown by Apt-Vv01. The invention alsoprovides a screening method for recognizing an aptamer of vibrio vulnificus. Property recognition and application are performed on the aptamer according to the affinity of the aptamer to the vibrio vulnificus. The aptamer consists of 149 bases and has a special secondary structure. The aptamer can be applied to rapid detection of vibrio vulnificus in medical diagnosis, environmental detection, food processing, aquaculture and the like. The aptamer effectively protects water bodies from being contaminated by the pathogen, and has wide application value especially in diagnosis, aquaculture andwater pollution prevention.

The invention discloses bacillus licheniformis HYT-9 with broad-spectrum bacteriostasis activity and a preparation method and application of a bacterial agent of the bacillus licheniformis HYT-9. Thebacillus licheniformis is named HYT-9, and the preservation number of the bacillus licheniformis is CGMCC NO. 17225. The bacteriostatic test shows proves that the bacillus licheniformis has significant bacteriostatic activity for multiple pathogenic bacteria, especially staphylococcus aureus, escherichia coli, streptococcus, vibrio alginolyticus, vibrio splendidus, pseudomonas and the like, and can be taken as a substitute product of antibiotics and applied to the fields of feed additives, antibacterial medicinal preparations, health care products, preservatives and the like. After fermentation in a tank of 15 L, the escherichia coli is taken as an indicator bacterium, doxycycline hydrochloride is taken as a contrast, and the bacteriostatic titer of fermented liquid of the bacillus licheniformis reaches 100 thousand ppm. The bacillus licheniformis has a wide market application prospect.

The invention relates to marine bacteria capable of antagonizing two important pathogenic vibrios of marine culture and an application of the marine bacteria. The antagonistic bacteria are marine-derived pseudoalteromonas agarivorans and are preserved in the common microorganism center of the China General Microbiological Culture Collection Center, and the preservation number of the antagonistic bacteria is CGMCC No17154. The pseudoalteromonas screened by the method is separated from Antarctic ice sea water, and the marine bacteria capable of antagonizing important pathogenic vibrio anguillarum and vibrio alginolyticus in aquaculture is a biological prevention and control potential strain with high prevention and control efficiency, wide prevention and control range and good environmentalsafety, and has good development and application prospects.

The invention relates to a primer probe composition capable of detecting five pathogenic bacteria at the same time. The primer probe composition comprises a sequence primer sequence or a probe sequence shown as SEQ ID No.1-15 as shown in a chart 1; a report radical group is modified at the 5' end of each probe sequence, and a quenching radial group is modified at the 3' end of each probe sequence; the report radial group has FAM, FITC, HEX, JOE and CY3, and the quenching radial group has TAMRA, BHQ and Eclipse. According to a detection method of five-fold real-time fluorescence quantification PCR, the five pathogenic bacteria such as salmonella, vibrio vulnificus, Shigella, staphylococcus aureus and Listeria monocytogenes can be detected at the same time in the same reaction system. Moreover, the five pathogenic bacteria are detected by using the five-fold real-time fluorescence quantification PCR method, the sensitivity and specificity of detection can be improved, the detection efficiency can be greatly improved, step complication and long detection period existing in conventional detection are remedied, and the primer probe composition cannot be applicable to the defects of large-scale quick detection.

The present invention relates to a dibenzoanthracene oxepanone compound from a marine fungus WH4-2 and a preparation method and application thereof. The marine fungus talaromyces stipitatus has the efficient V.parahemolyticus activity, the yield of an active substance, namely the dibenzoanthracene oxepanone compound containing an isoprene group is large, the dibenzoanthracene oxepanone compound isexpected to be applied to medicinal development of novel V.parahemolyticus, it is found that the compound further has Bacillus subtilis, Staphylococcus aureus ATCC29213 and Enterococcus faecium ATCC35667 resistance activity in further activity analysis, and the dibenzoanthracene oxepanone compound is expected to be developed into a broad-spectrum antibacterial medicine.