661 results about "Aflatoxins Toxicity" patented technology

The invention provides a hybridoma cell line 1C11 and an anti-aflatoxin general monoclonal antibody secreted by the same as well as the applications thereof. The hybridoma cell line 1C11 can be used for preparing a high-titer aflatoxin antibody, and a mouse hydroperitoneum antibody is measured to reach 5.12*106 by using an ELISA (Enzyme-Linked Immunosorbent Assay). The anti-aflatoxin general monoclonal antibody has high sensitivity, respectively reaches the IC50 (50% inhibiting concentration) of aflatoxin B1, B2, G1 and G2 to be 1.2, 1.3, 2.2 and 18.0 pg/mL, is the antibody with highest sensitivity among currently reported four aflatoxin antibodies, is used for measuring the total aflatoxin amounts, i.e. the total amounts of the aflatoxin B1, B2, G1 and G2 and has great practical application values.

The invention discloses a mycotoxin detoxification agent for feed and a preparation method and a feed additive thereof, and belongs to the field of feed additives. The mycotoxin detoxification agent can be used for detoxification of feed raw materials and compound feed of mycotoxin polluted corn, grains and the like. The mycotoxin detoxification agent consists of modified montmorillonite, yeast cell wall, chitosan and the like; and the preparation method has simple process. The novel mycotoxin detoxification agent for feed can effectively remove pollution of multiple mycotoxins such as aflatoxin, zearalenone, T2 toxin, vomitoxin and the like in the feed raw materials and the feed of the corn, the grains and the like, and prevent various diseases of livestock and poultry because of eating the mycotoxin polluted feed. The mycotoxin detoxification agent has simple preparation process, stable and controllable quality, convenience in use and low price, meanwhile has the advantages of supplementing nutrients, improving the disease resistance of cultured animal bodies and avoiding adsorbing nutrient elements in the feed, and guarantees the health and safety of the cultured animals.

The invention relates to an aflatoxin B1 flow lag immunization time distinguishing fluorescence rapid-detection kit and an application thereof. The kit comprises a fluorescent test strip and a sample reaction bottle containing an europium-labeled anti-aflatoxin B1 monoclonal antibody lyophilized product, wherein the fluorescent test strip comprises a cardboard, a water absorption pad, a detection pad and a sample pad are sequentially pasted on one surface of the cardboard from top to bottom, adjacent pads are connected at the connection in an overlapping manner, the detection pad treats a cellulose nitrate membrane as a base pad, a transverse quality control line and a detection line are arranged on the cellulose nitrate membrane from top to bottom, the quality control line is coated with a rabbit anti-mouse polyclonal antibody, and the detection line is coated with an aflatoxin B1 bovine serum albumin conjugate; and the anti-aflatoxin B1 monoclonal antibody is secreted by a hybridoma cell strain having a preservation number of CCTCC NO.C201015. The kit can be used for the quantitative determination of the content of the aflatoxin B1, and has the advantages of simple operation, rapidness and high accuracy.

The invention discloses a method for degrading aflatoxin, which makes use of gamma rays to irradiate a sample containing the aflatoxin, wherein the gamma rays are generated by a radioactive substance Co, and the irradiation dose is between 0 and 10 kGy not including 0, preferably between 2 and 10 kGy, more preferably between 4 and 10 kGy, particularly preferably between 6 and 10 kGy, and the most preferably 10 kGy. The sample containing the aflatoxin is a farm product containing the aflatoxin or a product obtained by processing the farm product, such as food, feedstuff, and the like. The method is particularly applicable to degrading the aflatoxin B1, not only can kill pathogenic microorganisms but also can degrade biotoxin therein, and does not generate any industrial pollution.

The invention discloses a complex microbial agent for producing soybean paste in Pixian County, which mainly comprises coculture of saccharomyces cerevisiae, candida utilis, lactobacillus plantarum and salt tolerant tetrads, and skim milk powder, glycerin, maltodextrin, trehalose and L-sodium glutamate, or mainly comprises coculture of saccharomyces cerevisiae, candida utilis, lactobacillus plantarum and salt tolerant tetrads, and porous starch, maltodextrin and polyvinyl pyrrolidone. A method for preparing the complex microbial agent for producing soybean paste in Pixian County comprises the following technological steps of: (1) activating and propagating strains; (2) inoculating; (3) co-culturing; and (4) adding a protective agent and drying. By using the complex microbial agent in the preparation of cooked soybean paste, the complex microbial agent maintains the characteristics of traditional soybean paste in Pixian County. Meanwhile, compared with the traditional technology, the complex microbial agent has the advantages of shortening the production period by 1/3, improving the amino nitrogen content by 3 to 8 times, improving the volatile aroma content by 2 to 4 times, and ensuring the aflatoxin B1 of 0 to 0.5ppm only.

The invention relates to a bacillussubtilis (Bacillussubtilis ANSB324 CGMCC No.3609) for decomposing aflatoxin, protein secreted by the same and an application thereof. The invention also provides a method for decomposing aflatoxin B1, G1 and M1 by using fermentation liquor of the strain or the protein secreted by the strain. By the method, after the bacillussubtilis and the aflatoxin B1 react for 48 hours, the decomposition rate of the B1 reaches 85%; after the bacillussubtilis and the aflatoxin G1 react for 72 hours, the decomposition rate of the G1 reaches about 66%; and after the bacillussubtilis and the aflatoxin M1 react for 72 hours, the decomposition rate of the M1 reaches about 68%. For the bacillussubtilis and the active protein secreted by the bacillussubtilis, the detoxifying activity is high, the specificity is strong, the action effect is mild, therefore, the bacillussubtilis provided by the invention is applicable to decomposing and removing the aflatoxin in feedstuff. Simultaneously, the bacillussubtilis provided by the invention has the advantages of strong bioactivity, good probiotics and stress resistance, and the like, and does not damage nutrient components in the feedstuff.

The present invention is fast detection method of aflatoxin B1 in food and feed. The fast detection method includes the following steps: adding ground sample to be tested in 2-10 g in test tube, adding methanol-sodium chloride aqua of 50-80 % concentration in 10-25 ml, ultrasonic extraction in water bath at 40-60 deg.c, and suction filtering to obtain filtrate in 4-10 ml; adding re-distilled petroleum ether in 2-4 ml into the filtrate, shaking, letting stand to laminate, taking the lower layer of solution in 1-4 ml and adding pure water in 4-20 ml; adding to micro immunoaffinity column of aflatoxin B1, washing with methanol aqua of 10-40 % concentration in 5-10 ml, eluting with methanol in 0.5-2.0 ml, collecting the eluted liquid, adding test agent A in 0.2-1.0 ml; and detecting in fluorescent spectrophotometry or similar instrument. The present invention has the features of simple process, high detection precision and stability, fast detection speed and high safety.

The invention provides an aflatoxin B1 (AFB1) magnetic separation enzyme-linked immunity quantitative detection method, belonging to the field of food safety immunoassay technique. The method adopts the immuno-detection principle of competition law; and AFB1 is connected with biological enzyme to prepare enzyme-labeled antigen reagent, anti-fluorescein isothiocyanate (FITC) antibody is absorbed onthe surfaces of magnetic particles to prepare magnetic separation reagent, and the FITC is connected with the AFB1 antibody to prepare anti-reagent. In a sample, the AFB1 competes with the enzyme-labeled AFB1 and is combined with a small amount of FITC-labeled anti-AFB1 antibody, so that antigen-antibody complex can be formed. After the magnetic separation reagent is added, the complex is caughtonto the surfaces of the magnetic particles by the anti-FITC antibody connected on the surfaces of the magnetic particles. After being washed, the product is finally added with substrate and detected.The method has the advantages that (1) the magnetic particles are used for replacing the traditional enzyme-labeled plate to be taken as a solid-phase carrier, so that immunoreaction is carried out under the approximate liquid phase condition; and the reaction is more complete and rapid, and has the characteristics of high specificity and good repeatability compared with the traditional enzyme-linked immuno sorbent assay (ELISA); furthermore, (2) by adopting one-step competition law principle, the time used for detection is short.

The invention relates to the field of bioengineering, in particular to a preparation method and an application of a microbial agent composition for enhancing secondary fermentation of a bean paste. The microbial agent composition consists of a saccharomycetes agent, an aspergillus oryzae agent and a pleospora agent; and the three microbial agents are prepared, and then the prepared microbial agents are mixed with the secondary-fermented bean paste, so as to promote the secondary fermentation of the bean paste. The whole process of the preparation method of the microbial agent composition provided by the invention is applicable to continuous industrial production; by-products (chili and radix platycodonis), from the production of Pixian bean paste, are comprehensively utilized; a production cycle can be shortened by 6 months, the content of amino nitrogen can be improved by 20% and the content of volatile aroma-producing components can be improved by more than 3 times (the contents of total ester, total acid and total aldehyde); by inoculating aflatoxin B1 with the saccharomycetes agent at a production peak (after being fermented for 30-60 days), an ester producing and aroma generating process can be enhanced, the metabolism of aspergillus flavus and partial aspergillus parasiticus can be competitively inhibited and the content of the aflatoxin B1 can be reduced, and the content of the aflatoxin B1 is lower than 0.5ppm, so that food safety is enhanced.

The present invention relates to a hybridoma cell line 3G1 and an anti-alfatoxin B1 monoclonal antibody produced by the hybridoma cell line 3G1. The hybridoma cell line 3G1 (CCTCCNO.C201014) can be used for preparation of a high titer anti-aflatoxin B1 monoclonal antibody, wherein an enzyme-linked immunosorbent assay (ELISA) method is adopted to determine a titer, and the titer is 6.40*10<6>. The anti-aflatoxin B1 monoclonal antibody of the present invention has characteristics of high sensitivity and good specificity, wherein 50% inhibiting concentration on aflatoxin B1 by the monoclonal antibody is 1.6 ng/mL, cross reaction rate with aflatoxin B2 is 6.4%, and cross reaction rates with aflatoxin G1 and G2 are less than 1%. In addition, the anti-aflatoxin B1 monoclonal antibody of the present invention can be used for determination of aflatoxin B1.

The invention belongs to the field of biological detection and provides a high-sensitivity immunochromatographic test strip for detecting total aflatoxin content quickly, which is characterized by comprising a test strip, wherein a water-absorbing pad, a detection pad, a gold label pad and a sample pad are adhered on one side of the test strip from top down; the pads are overlapped at the connected parts; the detection pad uses a nitrocellulose film as a substrate pad; transverse quality control lines and detection lines are arranged on the nitrocellulose film from top down; the detection lines are coated with aflatoxin B1-bovine serum albumin (AFB1-BSA) coupling and the quality control lines are coated with rabbit anti-mouse polyclonal antibody; and a nano gold-labeled anti-aflatoxin common monoclonal antibody, which is generated by hybrid tumor cell strain 1C11 having a collection number of CCTCC No.C201013, is sprayed on the gold label pad transversely. The test strip is used for detecting total aflatoxin content and has the characteristics of quick detection, simple operation and high flexibility.

The invention relates to a assaying method for the aflatoxin, specifically saying, it is a method of adopting immunity affinity column and fluorophotometer prepared by monoclonal antibody of aflatoxin to assayt the aflatoxin in the food, which belongs to biocytology and microbacteria toxic metabololic products assaying method technical field.The method is as following: first, preparing monoclonal antibody by the steps involving: immunizing mouse, cell fusion, screening crossing tumour and obtaining the cell line of the crossing tumourc, gathering monoclonal antibody, etc, then using the abovementioned monoclonal antibody to prepare the immunoaffinity column, last, after milling the food and feed, extracting the aflatoxin out of it with methanol/water solution, after filtering, letting the liquid sample flow through the affinity column to be refined, successively, eluting the AFT from the affinity column with methanol, and assaying the content of aflatoxin in the sample with a fluorophotometer.

The invention discloses a method for biodegrading mycotoxin in feed by using a mycotoxin degrading enzyme, and belongs to the field of feed resources. The method comprises the following steps: directly mixing degrading complex enzyme of aflatoxin, zearalenone and deoxynivalenol with moldy feed raw materials, concentrated feed or complete feed; and detoxifying the feed polluted by the mycotoxin to ensure that the degradation rate of the mycotoxin reaches over 80 percent. The method can effectively remove the mycotoxin from the feed, improves animal health and animal production performance, and can effectively solve the problem of feed resource shortage in China.

The invention discloses a preparation method for a mycotoxin adsorbent. According to the invention, the adsorbent having a full adsorption effect to multiple pathogenic mycotoxins is prepared according to the physicochemical property of mycotoxins, based on montmorillonite, and by means of the method of inorganic modification and organic compounding. The adsorbent can be added in feeds in a powder-shape mode according to a certain ratio for adsorbing common mycotoxins such as aflatoxin, ochracin, zearalenone and vomitoxin in feeds. According to the invention, the total adsorption amount of montmorillonite to mycotoxins is improved, and meanwhile the adsorbing property of montmorillonite to mycotoxins with various polarities is enhanced, so that the composite adsorbing effect is improved; and in addition, the adsorbent has the following characteristics of higher practicability, simple production technique, lower cost, and no toxicity or harmlessness.

A compact and portable analytical instrument dedicated to aflatoxin determination under minimum electrical power conditions employs a light emitting diode (LED) as light source with a peak output wavelength of 370 nm in addition to a 418 nm cut-off filter and a photodiode with a peak sensitivity of 140 nm. Thus, the relative amount of transmitted fluorescence energy at a wavelength of greater than 418 nm incident upon the aflatoxin is separated from the excitation light of 370 nm. In addition to the LED and the photodiode, the instrument preferably comprises amplifying, digital conversion, data storage, data transfer, display and power supply means and a graphical data output. The power supply regulator is a integrated circuit. As a result, current consumption is minimised, and thus battery life and instrument accuracy is maximised. The LED is powered by a constant current regulator, which minimises errors due to fluctuations in illumination intensity.

The invention aims at providing an immune magnetic particle for purifying aflatoxin samples and a preparing method and an application method thereof so as to solve technical problems in the prior art that aflatoxin sample purification and separation is complex in operation and low in separation efficiency and has large potential safety hazard. After aflatoxin antibodies and magnetic particles are reacted together for some time, the immune magnetic particle with specificity is prepared. After coarsely extracted aflatoxins and the immune magnetic particle are reacted together for some time, the aflatoxins are attached to the surface of the immune magnetic particle in affinity mode. After magnetic separation, the aflatoxin samples can be purified so that the relative pure aflatoxin samples can be obtained, and further the relative pure aflatoxin samples are used for fluophotometers, high performance liquid chromatographs (HPLCs) and enzyme-linked immuno sorbent assay (ELISA).

The invention discloses a bacillus amyloliquefacien for degrading aflatoxin B1 in peanut meal, and belongs to the technical field of applied microbiology. According to the invention, a bacillus amyloliquefacien capable of significantly inhibiting the growth of Aspergillus flavus and efficiently degrading aflatoxin B1 can be obtained by screening and the content of aflatoxin B1 in detoxified peanut meal is lower than the national limited standard and achieves the safe feeding level after the bacillus amyloliquefacien is applied to moldy peanut meal. Furthermore, the detoxification mechanism of the strain is the degradation effect of extracellular metabolites, and generation of the active substance is of a non-induced type, which is an inherent attribute of the strain. The features of the strain provide the possibility for biocontrol of Aspergillus flavus and aflatoxin B1 in industries such as feed and foods and the strain has very high application values.

The invention provides Lysobacter capable of efficiently degrading aflatoxin B1 and ochratoxin A and application of the Lysobacter and particularly provides double-function Lysobacter sp. CW239 and application thereof to the degradation of low-pollution concentration aflatoxin B1 and ochratoxin A. Compared with existing aflatoxin degrading bacteria, the Lysobacter sp. CW239 has the advantages that the Lysobacter sp. CW239 can achieve excellent degrading effect under a low-concentration toxin pollution condition; under a liquid fermentation condition, in fermentation broth, which contains the aflatoxin B1 and ochratoxin A, with the final concentration of 20 microgram/L, the 12-hour ochratoxin A degradation rate of the Lysobacter sp. CW239 is 53.1%, and the 48-hour degradation rate reaches 99.8%; the 12-hour aflatoxin B1 degradation rate of the Lysobacter sp. CW239 is 42.5%, and the 48-hour degradation rate reaches 83.4%; when the Lysobacter sp. CW239 is used for processing feed (with the final concentration of 20 microgram/kg) polluted by toxins, the 48-hour ochratoxin A degradation rate is 68.7%, and the 48-hour aflatoxin B1 degradation rate is 52.1%; the Lysobacter sp. CW239 has substantial application value and significance when being applied to food and feed bio-detoxification.

The present invention relates to preparation method of aflatoxin immuno-affinity column, including steps of: preparation of hydrazine antibody: employing sodium periodate oxidating unconjugated active area Fc hydroxoy of antibody to aldehyde group, and reacting with adipic dihydrazide to gain hydrazine antibody; preparation of aldehyde solid substrate: oxidating solid substrate containing continued diol structure by sodium periodate to gain aldehyde solid substrate; coupling of antibody and solid substrate: mixing hydrazine antibody and aldehyde solid substrate, adding sodium borohydride to seal unreacted aldehyde group on solid substrate after oscillating reaction to gain coupling antibody solid substrate; column packed. The present invention couples the unconjugated active area Fc end of antibody to solid substrate to remain activity of antibody better; hydrazide derived on antibody instead of solid substrate to avoid nonspecific adsorption of hydrazide group; the combined affinity column not introducing new groups ( such as carboxyl, amido and so on) except coupling antibody to avoid nonspecific adsorption from them.