81results about How to "No risk of poisoning" patented technology

The invention discloses a porcine streptococcus disease and haemophilus parasuis disease combined inactivate vaccine and a preparation method thereof. The preparation method comprises the following steps of: a, respectively carrying out enrichment culture on a porcine streptococcus strain, a haemophilus parasuis strain and a haemophilus parasuis strain to obtain a porcine streptococcus strain bacterial solution, a haemophilus parasuis strain bacterial solution and a haemophilus parasuis strain bacterial solution; b, respectively adding a formaldehyde solution into the porcine streptococcus strain bacterial solution, the haemophilus parasuis strain bacterial solution and the haemophilus parasuis strain bacterial solution, and inactivating; c, mixing the collected porcine streptococcus strain bacterial solution, the haemophilus parasuis strain bacterial solution and the haemophilus parasuis strain bacterial solution, adding Tween-80 for preparing a water phase, preparing white oil, Span-80 and aluminium stearate into an oil phase, mixing the water phase with the oil phase to prepare a uniform emulsion, i.e. an oil emulsion inactivating vaccine; and 4, sub-packaging the oil emulsion inactivating vaccine. The porcine streptococcus disease and haemophilus parasuis disease combined inactivate vaccine can effectively prevent the porcine streptococcus disease and haemophilus parasuis disease, does not have hidden danger of scattering viruses and is safe and reliable; and the immunization is realized by one vaccine, thus the cost is reduced.

An deactivated multi-bacterium vaccine for preventing the mastitis of milk cow is composed of Staphylococcus aureus, agalactic streptococcus, milk-stopping streptococcus, breast streptococcus, colibacillus and aluminum hydroxide gel. Its preparing process includes such steps as choosing the pathogenic bacterial strain of said mastitis, preparing seed liquid and reproduction liquid, purity checking, deactivating, preparing vaccine, and discriminating.

The invention relates to an activated toxic antibody kit for detecting a porcine reproductive and respiratory syndrome virus, which is characterized by being implemented by comprising the following steps of: after diluting a serum sample diluent to be detected twice, adding 100 microliters into an ELISA (Enzyme-Linked Immuno Sorbent Assay) plate for hatching at 37 DEG C for 1 hour; setting positive control, negative control and blank control; cleaning for five times with washing liquid every two minutes; then adding 100 microliters into each combined monoclonal antibody hole, hatching at 37 DEG C for 1 hour and cleaning for five times with washing liquid every two minutes; then adding goat-antimouse IgG-HRP ligature for acting at 37 DEG C for 1 hour and cleaning for five times with washing liquid every two minutes; then adding a developing substrate solution; mixing the substrate A with the substrate B according to the proportion of 1:1; adding 100 microliters into the ELISA hole; developing for 15 minutes by keeping in dark place; and adding 50 microliters of stop solution and detecting the OD490 value. The judging result is set according to the principle that the sample corresponding to the inhibition ratio PI which is equal to (OD uninhibited-OD sample)/OD uninhibited*100 percent, PI) 30 percent is positive, and the sample smaller than 30 percent is negative. The kit method has mature technology and strong repeatability, can effectively distinguish porcine reproductive and respiratory syndrome virus active toxic and inactivated toxic antibody and can be finished just by a common researcher.

The invention relates to a method for preparing a genetic engineering product, in particular to a prokaryotic expression protein of VP73 gene from African swine fever virus (ASFV) and a preparation method thereof. The preparation method comprises: artificially synthesizing the whole-length sequence of VP73 gene according to the sequence of the VP73 gene from ASFV in GenBank, constructing a recombinant expression vector pET32a-VP73, sequencing, verifying, transforming prokaryotic expression recipient bacteria E.coli BL21(DE3), and inducing expression by isopropyl-1-thio-beta-d-galactopyranoside (IPTG), wherein the molecular weight of the recombinant fusion protein is about 65KD. Protein purified by nickel column affinity chromatography can undergo a specific immune imprinting reaction with ASFV positive serum and avoid cross reaction with viruses such as swine fever virus, hog cholera virus, porcine circovirus, porcine reproductive and respiratory syndrome virus, swine influenza virus and pseudorabiesvirus. Experiments show the expressed protein has high detection sensitivity, and high specificity. When the antigen is used for detection, risk of spreading poison is avoided. And the antigen can be used as a detection antigen for use in an enzyme-linked immuno sorbent assay (ELISA) method for identifying an ASFV antibody.

The invention provides an inactivated vaccine for streptococcus suis and pasteurella multocida diseases and a preparation method thereof, which can prevent streptococcus suis infection caused by a plurality of different sero-group streptococcus and infection caused by a plurality of different capsular serotype pasteurella multocida, achieves multiple preventions with one injection clinically, and is lowered in cost, free from the hidden hazard of virus dispersion, safe and reliable. The invention further provides a method for preparing the inactivated vaccine for streptococcus suis and pasteurella multocida diseases.

The invention discloses a novel recombinant Akabane virus nucleotide capsid protein, gene encoding the recombinant protein and the process for preparing the recombinant protein. The recombinant protein has the amino acid sequence represented by SEQ ID No:1, the gene encoding the recombinant Akabane virus nucleotide capsid protein has the amino acid sequence represented by SEQ ID No:2. The preparing process of the recombinant protain comprises the following steps: constructing recombinant Akabane virus nucleotide capsid protein prokaryotic expression vectors, transforming bacillus coli with the recombinant Akabane virus nucleotide capsid protein prokaryotic expression vectors, inducing recombinant nucleotide capsid protein expression with IPTG, reclaiming and purifying the expressed recombinant nucleotide capsid protein.

The invention discloses an immunofluorescence reagent for detecting an E-type enterovirus and a detection kit thereof. A direct immunofluorescence detection method set up by applying the immunofluorescence reagent for detecting the E-type enterovirus can be used for clinically fast diagnosing E-type enterovirus infection and used for positioning and authenticating an E-type enterovirus antigen in tissue. The time for direct immunofluorescence detection is short, and a result can be reported within 40 min; specificity is high, and the immunofluorescence reagent only reacts with the E-type enterovirus; the immunofluorescence reagent is high in repeatability and accuracy, the coincidence rate between results of immunofluorescence reagent and results of an indirect immunofluorescence method is 90% or more, and the immunofluorescence reagent can be used for fast diagnosing E-type enterovirus infection.

The invention discloses a lucid ganoderma bag cultivation culture medium capable of increasing the content of ganoderan and selenium. The lucid ganoderma bag cultivation culture medium comprises, by weight, 75 parts of wood flour, 20 parts of wheat bran, 2 parts of corn flour, 1 part of lime, 1 part of gypsum and 1 part of sugar. A selenium-enriched nutrient solution is used for adjusting the ratio of the materials to water to be 1: (1.3-1.5). The selenium-enriched nutrient solution is obtained according to a preparation method including: (1), preparing 100 parts of purified water, 1.0 part of L-selenium methionine and 5 parts of corn flour; (2), mixing the raw materials in the step (1), putting the mixture into a high-pressure tank, increasing the pressure to 4kg/cm2 to even the pressure, keeping the pressure for 2-3 minutes, recovering the normal pressure within 1-3 seconds to obtain size, inoculating the size with mature liquid bacillus subtilis cultures by volume percent of 10% at the temperature of 25 DEG C, putting the size into a fermentation tank, injecting sterile air into the fermentation tank, stirring the fermentation tank at the speed of 250r/min, performing fermentation at the temperature of 35-37 DEG C for 4 hours, increasing the temperature to 70 DEG C quickly after fermentation, and keeping 1.0 hour of inactivation to obtain the lucid ganoderma bag cultivation culture medium.

The invention discloses a Ganoderma lucidum karst cultivation method for improving ganoderma lucidum polysaccharides and selenium content. The method comprises the steps of 1, preparation of a culture medium, wherein the culture medium is prepared from, by weight, 75 parts of sawdust, 20 parts of wheat bran, 2 parts of corn flour, 1 part of lime, 1 part of gypsum and 1 part of sugar, and the ratio of the materials to water is adjusted to 1:(1.3-1.5) with a selenium-rich nutrient solution; 2, material mixing, bagging, sterilization and inoculation; 3, spawn running, wherein the inoculated culture medium is placed in a greenhouse for spawn running, at the beginning, the greenhouse temperature is controlled to be 26-28 DEG C, at the 20th day after inoculation, the greenhouse temperature is increased to be 34 DEG C and kept for 20-24 h, and then recovered to be 26-28 DEG C; 4, ganoderma lucidum karst germination, wherein after hyphae grow to be filled with the bag, ganoderma lucidum karst enters into a ganoderma lucidum karst germination period, and the greenhouse temperature is increased to be 34 DEG C at the 5th day, the 10th day, the 15th day and the 20th day in the ganoderma lucidum karst germination period respectively, kept for 20-24 h and then recovered to be 26-28 DEG C.

The invention provides a gB subunit recombinant protein of a porcine pseudorabies virus, and a preparation method and application of the gB subunit recombinant protein. The preparation method comprises the following steps of 1) cloning an optimized gB gene sequence into an eukaryotic expression vector to obtain a recombinant plasmid containing a gB subunit protein coding gene of the pseudorabies virus; 2) transfecting the recombinant plasmid containing the gB subunit protein coding gene of the pseudorabies virus into an expression cell; 3) culturing, screening and domesticating the expressioncell in the step 2) to obtain a highly expressed cell strain; and 4) fermenting and culturing the cell strain in the step 3), and performing purification to obtain the gB subunit protein of the pseudorabies virus. The invention provides the gB subunit protein of the porcine pseudorabies virus, and the gB subunit protein can be industrially produced in a large scale; the preparation method is simple; the cost is low; and the yield is much higher than that of an existing baculovirus expression system.

The invention discloses a double-antibody sandwich ELISA kit for detecting an E-type enterovirus pathogen. VP1 protein expressed by a VP1 gene with the base sequence shown as a sequence table SEQ ID NO.1 is adopted to immunize BALB/C mice to prepare a polyclonal antibody serving as a capture antibody, VP2 protein expressed by a VP2 gene with the base sequence shown as a sequence table SEQ ID NO.3 is adopted to immunize BALB/C mice to prepare a monoclonal antibody, and horse radish peroxidase (HRP) is marked. Safety is high, and the coincidence rate between kit detection results and RT-PCR detection results is 100%. The kit can be stored for 6 months at the temperature of 4 DEG C, and no obvious difference exists between detection results and those of a conventional method.

The invention provides a clostridium perfringen alpha toxin genetic engineering vaccine and a preparation method thereof. Compared with wild type alpha toxins, clostridium perfringen alpha toxin recombination protein is characterized in that the 176th site histidine of an amino acid sequence is mutated to obtain asparagine. The clostridium perfringen alpha toxin genetic engineering vaccine is prepared from detoxifcation clostridium perfringen alpha toxin recombination protein of self-induction secretory expression. The vaccine has the advantages of being safer, better in immune efficacy, simpler in technology, lower in cost and the like, and can effectively solve the problems that in the prior art, the production technology of the vaccine is complexer, and the cost is higher.

The invention discloses a method for planting selenium-rich peanuts. The method comprises the following steps of 1, soil preparation and fertilization; 2, seed selecting and sowing; 3, field management, and further comprises the step that in the peanut blooming acicula forming period, the fruiting period and the later growth and development period, a cultivation nutrient solution is spayed to leaf surfaces. The cultivation nutrient solution is prepared through the following method which includes the steps that 1, raw materials are prepared, wherein 100 parts of pure water, 0.6-1.0 part of L-selenium methionine, 1.5-2.5 parts of water-soluble compound fertilizer with NPK being 15-15-15 and 0.1-0.3 part of ethoxy fibers are prepared; 2, the raw materials in the step 1 are mixed and then put in a high-pressure tank, the pressure is increased to 4 kg/cm<2>, the pressure is made to be even and kept for 2 min to 3 min, then normal pressure is restored within 1 s to 3 s, pulp is obtained, the pulp is inoculated with bacillus subtilis liquid strains cultured to be mature at the temperature of 25 DEG C according to the volume percent of 5%-10%, the materials are put in a fermentation tank, sterile air is introduced into the fermentation tank, stirring and fermentation are carried out, after fermentation is completed, the temperature is quickly raised to 70 DEG C to be kept for 0.5 h to 1 h, then inactivation is carried out, and the selenium-rich peanuts are obtained.

The invention discloses a selenium-enrichment rice cultivation technology. The technology is implemented as follows: a nutrient solution is sprayed at the last stage of rice tillering, wherein leaf surfaces need to be covered uniformly by the solution. The nutrient solution is prepared as follows: (1), raw materials are prepared; to be specific, 100 parts of purified water, 1.5 to 2.5 parts of L-selenomethionine, 2.5 to 3.5 parts of water-soluble compound fertilizer with the NPK proportion of 15 to 15 to 15, and 0.5 to 1.0 part of hydroxyethyl cellulose; (2), after mixing of the raw materials prepared at the step (1), the mixed raw materials are placed into a pressure tank, pressurization is carried out to reach 4kg/cm<2> and the pressure is kept to be uniform for 2 to 3min, and then the pressure returns to a normal pressure within 1 to 3s to obtain serous fluid; liquid bacillus subtilis spawn with maturation is introduced into the serous fluid based on a percent by volume of 5 to 10% at a temperature of 25 DEG C, the mixture is placed into a fermentation cylinder, sterile air is guided into the fermentation cylinder, stirring and fermentation are carried out; and after fermentation completion, the temperature rises to 70 DEG C rapidly and the temperature is kept for 0.5 to 1.0h for inactivation, thereby obtaining the nutrient solution.

The invention discloses a method for cultivating selenium-rich peanuts. The method comprises the following steps of 1, soil preparation and fertilization; 2, seed selecting and sowing; 3, field management and further comprises the step that a cultivation nutrient solution is sprayed to leaves in the peanut blooming and seed growing period, the fruiting period and the later growth and development period. The cultivation nutrient solution is prepared through a method including the following steps that 1, raw materials are prepared, wherein 100 parts of purified water, 0.6-1.0 part of L-selenomethionine, 1.5-2.5 parts of water-soluble compound fertilizer with NPK being 15-15-15 and 0.1-0.3 part of hydroxyethyl cellulose are prepared; 2, after the raw materials in the step 1 are mixed, the mixture is placed in a steam cavity charge container, the pressure is 4 MPa, the mixture is heated to 150 DEG C, the temperature is kept for 2 min, then the pressure is fast released within 1 s to 3 s, slurry is obtained and inoculated with 5%-10% by volume of bacillus subtilis liquid strain cultured to be mature at the temperature of 25 DEG C, the mixture is placed in a fermentation tank, sterile air is introduced into the fermentation tank, stirring fermentation is carried out, after fermentation is completed, the temperature is fast raised to 70 DEG C to be kept for 0.5-1.0 h, inactivation is carried out, and the cultivation nutrient solution is obtained.

The invention discloses a formulated fertilizer for double-season Sophora japonica. The formulated fertilizer is prepared according to a method including the steps of 1), preparing raw materials, namely preparing 100 parts of purified water, 2.0-4.0 parts of L-selenomethionine,10-15 parts of water-soluble compound fertilizer with NPK (nitrogen, phosphorus and potassium) in a ratio of 15 to 10 to 20, 1.0-2.0 parts of hydroxyethyl cellulose and 200 parts of decomposed farmyard manure; 2), mixing all the raw materials except for the decomposed farmyard manure, putting a mixture into a steam explosion pot, heating to 150 DEG C at the pressure of 4 MPa, holding for 2 minutes, rapidly releasing pressure within 1-3 seconds to obtain slurry, inoculating a matured bacillus subtilis liquid strain according to a percent of 5-10% by volume at 25 DEG C, disposing the matured bacillus subtilis liquid strain into a fermentation tank, feeding sterile air into the fermentation tank with stirring fermentation, rapidly increasing the temperature to 70 DEG C after fermentation, holding the temperature for 0.5-1.0 hour to inactivate, and mixing with the decomposed farmyard manure so as to obtain the formulated fertilizer.

The invention discloses a truncated recombinant protein of a Riemerella anatipestifer (RA) serum type 1 siderophore receptor protein (SRP) and application of the truncated recombinant protein. The SRP truncated recombinant protein can generate specific reaction with a serum type 1 RA clonal antibody, and can be used as an ideal antigen for detecting a serum type 1 RA antibody. The truncated recombinant protein of the RA serum type 1 SRP, prepared by the invention, is not all bacteria, and has no danger of dispersing toxicity; in addition, the solubility of the protein during prokaryotic expression is increased, and the large-scale production cost of downstream can be effectively reduced; the truncated recombinant protein is suitable for industrial large-scale production and facilitates the research on laboratory vaccines.