100 results about "Streptococcus mastitidis" patented technology

The invention provides proteins from group B streptococcus (Streptococcus agalactiae) and group A streptococcus (Streptococcus pyogenes), including amino acid sequences and the corresponding nucleotide sequences. Data are given to show that the proteins are useful antigens for vaccines, immunogenic compositions, and/or diagnostics. The proteins are also targets for antibiotics.

The invention relates to an aquaculture animal disease control technology and in particular relates to a tilapia mossambica source streptococcus agalactiae low virulent strain and application thereof. The tilapia mossambica source streptococcus agalactiae low virulent strain is YM001 and is preserved in CCTCC (China Center For Type Culture Collection) with the preservation number of CCTCCM2014045. The tilapia mossambica source streptococcus agalactiae low virulent strain is subjected to activation culture and fermentation culture, bacterial cells of a 106-109CFU/mL low virulent strain YM001 and a sterile phosphate buffered solution are mixed as a low virulent live vaccine used for preventing and controlling a tilapia mossambica streptococcus disease, the injection medication concentration for each tilapia mossambica is 105-108CFU, the oral medication concentration for each tilapia mossambica is 108-109CFU, and the soaking medication concentration is 107-108CFU/mL. The invention relates to strain identification, toxicity test, toxicity return safety and stability, immunogenicity and immune protection effect of a low virulent strain, and the tilapia mossambica streptococcus disease vaccine prepared by adopting the low virulent strain is good in immune protection effect and strong in safety and stability.

The invention discloses a method for constructing an anti-mammitis transgenic mouse model and a special vector for the method. An anti-mammitis-related protein composition comprises lysostaphin and peptidoglycan endolysin B30; the peptidoglycan endolysin B30 derives from a Streptococcus agalactiae phage B30, and an amino acid sequence of the peptidoglycan endolysin B30 is sequence 2 in a sequence table; and an amino acid sequence of the lysostaphin is sequence 3 in the sequence table. The experiment proves that a transgenic plasmid for mammary gland-specific expression of the lysostaphin and the peptidoglycan endolysin B30 is constructed by using a mammary gland-specific expression vector; and the anti-mammitis transgenic mouse model is constructed by a microinjection method, a transgenic mouse with an anti-bovine mastitis gene is obtained, and a technical model is provided for further producing anti-mammitis transgenic calves.

The invention discloses new pleuromutilin derivatives which have a structural formula (I) or a structural formula (II) shown in the specification, wherein in the formula (I), when n=0, R1 is Cl, CH3, OCH3 or NH2; and when n=1, R1=H, and R2=NH2. The compounds have favorable inhibiting action on Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Streptococcus mastitidis; the antibacterial activity of one compound having a phenyl group with ortho-substituting groups or para-substituting groups (such as Cl, CH3, OCH3 or NH2) is superior to the antibacterial activity of one compound having a phenyl group with meta-substituting groups; and part of compounds having para-substituting aryl groups or ortho-substituting aryl groups are the same with valnemulin in the antibacterial activity on Staphylococcus epidermidis or Streptococcus mastitidis, and can be used for the preparation of antibacterial drugs. The synthesis method of the compounds has the advantages that the raw materials are accessible, the price is low, the operation process is simple, the products are easy to separate and purify, the yield is high and the total yield is 35-45%.

The invention relates to bacillus cereus NY5 capable of effectively inhibiting tilapia-source streptococcus agalactiae. The strain is preserved in the China General Microbiological Culture Collection Center (CGMCC) and the preservation number is CGMCC No.9372. The bacillus cereus is aerobic bacteria and can effectively inhibit growth of tilapia-source streptococcus agalactiae. Through a nitrate nitrogen experiment, removal efficiency can reach 100%. The strain also has functions of casein hydrolysate and starch and has passed safety detection. In a water body with concentration of bacillus cereus NY5 being 2*107cfu/ml and under the condition of injecting 100 microliters of a NY5 bacterium solution at the concentration of 2.1*106cfu/ml, no death or other abnormal phenomena happen to tilapia. According to the invention, a useful strain source is provided for effectively inhibiting growth of tilapia-source streptococcus agalactiae and efficiently degrading nitrite nitrogen, protolysate and starch under the aerobic condition, and the application range of bacillus cereus in the aspect of functions is widened. The bacillus cereus NY5 has high application value.

The invention discloses a preparation method and application of a tilapia source streptococcus agalactiae recombinant GroEL protein vaccine. The recombinant GroEL protein vaccine is prepared by tilapia source streptococcus agalactiae recombinant GroEL protein. An amino acid sequence of the tilapia source streptococcus agalactiae recombinant GroEL protein is as shown in SEQ ID NO.2. In the invention, tilapia source streptococcus agalactiae is taken as a research object, a GroEL gene is obtained from a tilapia source streptococcus agalactiae genome by cloning, the cloned GroEL gene is connected with a pET28a(+) carrier to construct an expression vector, and the vector is subjected to prokaryotic expression and purification in E.coli BL21(DE3) to obtain the recombinant GroEL protein vaccine with strong specificity and an immune protective efficacy of 68.61+/-7.39 percent, so that a foundation is laid for immune prevention and treatment of a tilapia streptococcus agalactiae disease.

This invention relates to new vaccines against microorganisms based on antigenically mimetic peptides. The invention also relates to methods of discovering such mimetic peptides by first screening peptide-display phage libraries with antibodies against the microbial carbohydrates(s) of interest to locate antigenically mimetic peptides. Vaccines against Group B Streptococcus, or Streptococcus Agalactiae, are preferably produced using this method.

The invention discloses recombinant lactococcus lactis based on the fbsA gene of Streptococcus agalactiae. The recombinant lactococcus lactis comprises the fbsA gene of Streptococcus agalactiae or segments of the fbsA gene, the fbsA gene or the segments of the fbsA gene preferably has (or have) a sequence shown in SEQ ID No. 1, the deposit number of the recombinant lactococcus lactis is CCTCC M 2020140, and expression of recombinant FbsA protein with a theoretical molecular weight of 34.6 kDa can be induced under the culture condition that niusin exists. The invention also provides a vaccine for the Streptococcus agalactiae disease of Oreochromis sp., and the vaccine is prepared from the recombinant lactococcus lactis, and is capable of producing protective antibodies in Oreochromis sp. after Oreochromis sp. is fed with the vaccine in an immune mode of oral gavage, so that Oreochromis sp. is assisted to resist infection of Streptococcus agalactiae effectively.

The invention discloses a rapid separation and identification kit for streptococcus agalactiae. In the kit, the following reagents are included: sheep defibering blood plate, 3% H2O2, staphylococcus aureus strains, a specific PCR (polymerase chain reaction) solution, and positive control DNA (deoxyribonucleic acid). The rapid separation and identification kit for streptococcus agalactiae provided by the invention has the following beneficial effects that bacterial morphological observation, catalase test, CAMP (cyclic adenylic acid) reaction and PCR detection are used together, so that identification procedures can be simplified, the specificity is strong, and the sensitivity is increased by more than 15%; and moreover, the identification cost is low, the time consumption is short (only needing 3 days), and streptococcus agalactiae strains can be obtained by separation and accurately identified. The kit can be used for separation and identification for the samples of the milk samples of human or dairy cows, cervical mucus, vaginal swabs and the like.

The present invention relates to polypeptides, epitopes and antibodies directed to these epitopes, more particularly to the Sip polypeptide of Group B Streptococcus (GBS), also called Streptococcus agalactiae which may be used to prevent, diagnose and/or treat streptococcal infection.

The invention provides a LAMP (Loop-Mediated Isothermal Amplification) primer group, a micro-fluidic chip and a kit for detecting reproductive tract pathogenic microorganisms, and belongs to the technical field of reproductive tract infection detection. The LAMP primer group comprises a streptococcus agalactiae detection primer group, an enterococcus faecalis primer group, a gardnerella vaginosisprimer group, a Candida albicans primer group and a Chlamydia trachomatis primer group. The above LAMP primer group is fixed on the micro-fluidic chip. The LAMP primer group, the micro-fluidic chip and the kit can quickly, sensitively and accurately detect streptococcus agalactiae, enterococcus faecalis, gardnerella vaginosis, Candida albicans and Chlamydia trachomatis by high repeatability.

The invention discloses a method for constructing a high-yield glutathione recombinant strain and application, and belongs to the field of bioengineering. The method for constructing the high-yield glutathione recombinant strain is characterized in that corynebacterium glutamicum for synthesizing a large amount of glutamic acid is used as an original strain, and a gene gshF from streptococcus agalactiae or genes gshA and gshB from escherichia coli are introduced to endow glutathione with the synthesis capability; further genome is subjected to deletion of genes aceD, mcbR and sdaA; a truncatedgene serA, a gene pgk, genes cysE and cysK derived from the escherichia coli and a gene cysE derived from arabidopsis thaliana are expressed at the same time, and a cysteine pathway is strengthened;and as the corynebacterium glutamicum can synthesize a large amount of the glutamic acid and the cysteine pathway is strengthened, the glutathione can be highly produced only by adding glycine in a fermentation process of obtained engineering bacteria.

The invention relates to a culture-independent semiquantitative detection kit of tilapia streptococcus agalactiae, which consists of a reagent A, a reagent B, a reagent C, a reagent D, a reagent E, a product specification and a test result record card. The kit can be used for directly detecting whether blood of tilapia carries the streptococcus agalactiae without pure culture by a method provided by the invention, and semiquantitatively detect the quantity of the streptococcus agalactiae in the blood.

The invention discloses streptococcus agalactiae BibA recombinant protein, as well as a coding gene, a preparation method and an application thereof. The streptococcus agalactiae BibA recombinant protein provided by the invention is a protein sequence as shown in sequence No. 2. The invention also provides the coding gene, the preparation method and application of the protein. The streptococcus agalactiae BibA recombinant protein has the beneficial effects that streptococcus agalactiae is used as a research object, streptococcus agalactiae DNA is cloned to obtain a BibA gene, the cloning and sequencing of the gene sequence are completed, the prokaryotic expression and immunizing protection of the streptococcus agalactiae are researched, a vaccine development candidate antigen with high specificity and immune protection force is obtained, and the foundation is laid for prevention of streptococcus agalactiae diseases. The streptococcus agalactiae BibA recombinant protein has the advantages that the streptococcus agalactiae BibA gene is obtained by cloning, BibA recombinant expression plasmids are constructed, the BibA recombinant protein is obtained, westernblot detection results show that the BibA recombinant protein has relatively good reactogenicity, and animal (rat) experiments prove that the recombinant protein has certain protectiveness.

The invention relates to an aquaculture animal vaccine, in particular to an attenuated live vaccine prepared from a tilapia mossambica streptococcus agalactiae natural attenuated strain TFJ0901. The streptococcus agalactiae natural attenuated strain TFJ0901 is preserved in China Center for Type Culture Collection of Wuhan University, China, and the preservation number is CCTCC NO: M 2016792. The ingredients of the attenuated live vaccine are live cells of the natural attenuated strain TFJ0901 and a phosphate buffer solution, and the cell concentration is 10<6>-10<8> CFU/ml. The vaccine has high safety and immunogenicity and can effectively prevent tilapia mossambica streptococcus agalactiae.

The invention relates to recombinant Lactococcus lactis comprising a fusion gene of a sip gene and a pgk gene of Streptococcus agalactiae, and the fusion gene is introduced into the Lactococcus lactisby a plasmid vector. The fusion gene has a DNA sequence as shown in SEQ No. 1. The plasmid vector is a pNZ8148 vector. The lactococcus lactis is Lactococcus. lactis NZ9000. The invention further provides a tilapia mossambica streptococcus agalactiae disease vaccine. The tilapia mossambica streptococcus agalactiae disease vaccine comprises the recombinant lactococcus lactis. Wherein the recombinant lactococcus lactis is the recombinant lactococcus lactis induced by nisin. The vaccine is the vaccine administrated by oral administration and intragastric administration. The tilapia mossambica streptococcus agalactiae disease vaccine comprises the recombinant lactococcus lactis with the concentration of 2*10 <10> cfu mL <-1>, and the administration dosage is 200 [mu]L.

The invention discloses a preparation method and application of a micro-capsule coated egg yolk antibody IgY resistant to main pathogenic bacteria of dairy cow mastitis. A compound mycoprotein antigen is prepared by selecting streptococcus agalactiae, streptococcus dysgalactiae, staphylococcus aureus and escherichia coli and is mixed with an equal quantity of freund's adjuvant to prepare a multivalence antigen immune complex, laying hens are immune by adopting repeatedly alternative immune procedures and a muscular and subcutaneous multipoint injection means, a specific yolk immunoglobulin IgY resistant to the main pathogenic bacteria of dairy cow mastitis, namely DP-BM-IgY, is separated and purified from collected egg yolk, then homogeneous emulsification is conducted on the specific yolk immunoglobulin and a sodium alginate water solution and an emulsifying agent soya bean salad oil containing span-80, the emulsified solution is dropwise added into an encystation solution prepared from CaCl2 and chitosan for coating and curing, and the coated micro-capsules can effectively protect the activity of the DP-BM-IgY and have enteric solubility. The preparation method utilizes a micro-capsule technology and selects the chitosan and sodium alginate as coating materials (natural polysaccharides), and the egg yolk antibody is internally taken and free of any toxic or side effect.

The invention belongs to the technical field of aquaculture preparations, and discloses a feed additive for preventing streptococcus agalactiae, being prepared from the following components by mass percent: 10-15 percent of coptis chinensis, 7-15 percent of honeysuckle, 7-15 percent of isatis root, 10-20 percent of Houttuynia cordata, 20-40 percent of black soya bean and 0-20 percent of dendrobe. The additive is added into tilapia feed, has a highest immune protective rate of 95.21 percent for tilapia, and has practical and wide application values; the survival rate of the tilapia is 93.67 percent.

The invention relates to a tilapia streptococcus agalactiae vaccine and a preparation method thereof. According to bioinformatics principles, a part of polypeptide sequences of a surface anchoring protein of streptococcus agalactiae are analyzed and screened as an antigen for preparing the vaccine, in addition, a carbon nanotube carried surface protein system is constructed by using a chemical synthesis technology, and the selected polypeptide antigen is in nano combination with functionalized single-wall carbon nanotubes, so that a tilapia streptococcus agalactiae subunit vaccine is prepared.The polypeptide antigen screened by the invention is capable of remarkably increasing the antibody level of inoculated tilapia when being compared with an inactivated vaccine, and is capable of resisting attack of streptococcus agalactiae of high toxicity. In addition, the polypeptide antigen screened by the invention can be tightly and uniformly connected with surfaces of functionalization modified carbon nanotubes, and compared with independent polypeptide antigen groups, the polypeptide antigen is capable of remarkably improving the immunity of the tilapia.

The invention discloses a sophora flavescens flavonoid active fraction with tyrosinase inhibitory activity and antibacterial activity and a preparation method of the sophora flavescens flavonoid active fraction. The preparation method comprises the following steps: taking sophora flavescens medicinal residues as raw materials, changing wastes into valuables, adding ethanol for reflux extraction, filtering, drying filtrate to obtain an ethanol extract, dissolving the extract with methanol, loading to AB-8 type macroporous adsorbent resin, then performing gradient elution with ethanol with the concentrations of 10 percent, 20 percent, 30 percent, 40 percent, 50 percent, 60 percent, 70 percent and 80 percent, and collecting 50-70 percent of elution part. The active fraction prepared with thepreparation method disclosed by the invention has obvious inhibitory activity to tyrosinase, has an obvious antibacterial effect on staphylococcus aureus and streptococcus agalactiae, can also well prevent and treat a human pigmentation disease caused by excessive melanin and has obvious and extensive biological activity.

The invention discloses a method for weakening the toxicity of tilapia sourced streptococcus agalactiae. The method disclosed by the invention comprises the following steps of carrying out streak culture of a separately stored wild strain on a blood plate, and picking single colonies to a TSB culture medium to carry out shake culture at 25 DEG C; step 1, after culturing for 6 h, inoculating single colonies on a novel TSB culture medium to carry out shake culture at 25 DEG C, and subcultring every 6 h till 20 generations; step 2, inoculating the 20 generation of culture on a novel LTSB culture medium to carry out shake culture at 35 DEG C, and subcultring every 6 h till 40 generations; processing the 41 generation of culture by repeating the step 1 till processing the 60 generation of culture, and processing the 61 generation of culture by repeating the step 2 till processing the 80 generation of culture, wherein subcultring is continuously carried out according to the method; the toxicity of the strain is tested in the subcultring process, and subcultring is carried out till the toxicity of the strain is weakened. The method for weakening the toxicity of tilapia sourced streptococcus agalactiae is simple and rapid; the method for weakening the toxicity of a tilapia sourced streptococcus agalactiae wild strain is weak in toxicity and difficultly becomes toxic; the method has important significance and application value for developing attenuated vaccines of tilapia streptococcicosis.

The invention discloses a double-antibody sandwich enzyme-linked immunosorbent assay (double-antibody sandwich ELISA) kit for detection of Streptococcus agalactiae. The kit is composed of a perforated plate coated with 3A9 monoclonal antibody, a horse radish peroxidase labeled monoclonal antibody 4C12 and a chromogenic substrate 3'.3'.5,5'-tetramethyl benzidine (TMB). The preparation method of the kit includes the steps of: (1) preparing the perforated plate coated with 3A9 monoclonal antibody; (2) preparing the horse radish peroxidase labeled monoclonal antibody 4C12; and (3) preparing the chromogenic substrate 3'.3'.5,5'-tetramethyl benzidine.