34 results about "Locoweed" patented technology

A detoxicating agent for locoweed contains proportionally medical ferrous salt, medical zinc salt and medical copper salt. A raw material used for preparing ensilage contains said detoxicating agent (0.8-1.2 Wt. Portions), locoweed (800-1200), and additive (16-32) or grass (1200-2700).

This invention discloses a novel method for extracting swainsonine from locoweed. The method comprises: (1) extracting locoweed powder with ethanol at a ratio of 1:(6-10) for 3 times (4 h each time), filtering, recovering ethanol, dissolving the residue with 3-5 times of ethanol, recovering ethanol to obtain an extractum, adding 2% acetic acid to adjust the pH value to 4.5-6, adding 50-100 times of chloroform, and recovering the chloroform layer; (2) extracting the acidic water layer with 50 times of chloroform for twice, recovering chloroform, drying the acidic water at 50-60 deg.C to obtain crude alkaloids, completely dissolving in 1 M ammonium solution, adding 10% NaOH solution (1 mL/45-75 g) and methanol (5 mL/45-75 g) into alkaloids solution, extracting with dichloromethane repeatedly until the solution color is light, adding ammonium solution to completely dissolve the residue, extracting with dichloromethane, and recovering dichloromethane to obtain crude swainsonine; (3) dissolving with methanol, evaporating methanol, and sublimating to obtain white needle-like crystals. The method has such advantages as easy operation, and high extraction rate.

Belonging to the field of chemical analyses on natural products, the present invention relates to a method for extracting, separating, purifying and testing natural products, in particular to a technique for extracting swainsonine by continuous column chromatography. The present invention resolves the defects of the prior art, including a complex process, high cost and low yield rate. The solution orderly includes the following steps: (1) the preprocessing of locoweed; (2) the acquisition of locoweed extract; (3) the preparation of a crude product: The extract is roughly separated by polar macroporous adsorption resin and, after TLC test, merged with eluant containing swainsonine to serve as the crude product for later use; (4) the purification of the swainsonine: The crude product is purified by silica gel column chromatography, and by the TLC test and the GC test, the parts of the swainsonine, which have relative content larger than or equal to 10 percent, are respectively collected; (5) the acquisition of a pure swainsonine product by the HPLC test; the parts collected in the step (4) undergo the HPLC test in order to collect eluate in the appearance time period of the swainsonine, and after the solvent is evaporated, the pure swainsonine product can be produced by recrystallization.

The invention relates to a method for directional separation and authentication of swainsonine fungal endophyte Undifilum oxytropis produced through oxytropis kansuensis. When the fungal endophyte Undifilum oxytropis is separated from oxytropis kansuensis, bacterial strains of other species can be inevitably separated, and therefore separation efficiency is affected. Stems and mature seeds of oxytropis kansuensis are sequentially cleaned through distilled water, ethanol, sterile water and sodium hypochlorite, tissue is sheared into small blocks and placed on a PDA flat plate, the flat plate is filled with CO2 and sealed by a fresh-keeping film, the tissue is cultured at indoor temperature under the illumination condition, after mycelia grow on incisions, the mycelia at the top ends are picked and transferred to a culture medium for purification, the mycelia are scraped, and then the bacterial strains are inoculated to a test tube slant for refrigeration; genome DNA of the separated bacterial strains is extracted, and sequence alignment is conducted in a Gene Bank. Only one fungus is separated from locoweed, interference of fungal endophyte of other species is reduced, the method is easy and convenient to implement and feasible, and in the storage process, the bacterial strains are not prone to mutation or activity of the bacterial strains is not reduced.

The invention discloses a swainsonine degradation bacteria acetic acid calcium acinetobacter YLZZ-1, which is of short rod shape, 0.2 to 0.3Mum x 0.5 to 1.0Mum, gram-negative without capsule, germ andmotion; bacterial colony is circular, ivory-white, smooth on the surface, wet, elevated and regular in edge and not transparent; bacterial strain is artificially separated and screened out from locoweed growing soil in areas such as Ganshu, Tibet, Inner Mongolia and Qinghai and is preserved at the preservation center for typical culturing in China on Jul., 20, 2007 with preservation No. of CCTCCNO: M 207108. The invention has the advantages of degrading swainsonine which is main toxin of locoweed (Swainsonine, SW).

The invention relates to a traditional Chinese medicine compound pill used for preventing locoweed poisoning of animals. The traditional Chinese medicine compound pill is characterized by comprising the following raw materials by weight: 9 to 18 parts of dried orange peel, 9 to 18 parts of largetrifoliolious bugbane rhizome, 15 to 30 parts of honey-fried licorice root, 9 to 18 parts of tangshen, 9 to 18 parts of Chinese angelica, 9 to 18 parts of Chinese thorowax root, 30 to 60 parts of processed milkvetch root, 9 to 18 parts of bighead atractylodes rhizome and 5 to 10 parts of a trace element mixture. The traditional Chinese medicine compound pill provided by the invention is targeted at prevention of locoweed poisoning, has good security and enables breeding cost to be saved.

The invention relates to a method for detecting the content of swainsonine in locoweed endophytic fungi. The method comprises the steps: pre-treating an endophytic fungi hyphae sample to prepare a test solution; using a high performance liquid chromatograph evaporative light-scattering detector to detect the swainsonine in the locoweed endophytic fungi; taking a mixed solution of acetonitrile andammonium acetate buffer solution as a mobile phase, wherein the volume ratio of acetonitrile to ammonium acetate is 0.2-2 to 0.2-2; separating the swainsonine from other materials in the test sample by an HILIC (Hydrophilic Interaction Chromatography) column; and constructing a map by logarithm of swainsonine in different concentrations and logarithm of a corresponding response peak area so as toobtain a standard curve of the swainsonine. According to the method, the swainsonine in the endophytic fungi Undifilum oxytropis can be effectively separated, and the method can be directly applied to the quantitative analysis of the swainsonine. The method is good in separation effect, strong in specificity and high in accuracy, and the analysis method is reliable.

The invention discloses a traditional Chinese medicine compound preparation for treating locoweed poisoning of animals. The traditional Chinese medicine compound preparation is prepared from the following raw materials in parts by weight: 9-18 parts of elecampane, 9-18 parts of angelica sinensis, 15-30 parts of ligusticum wallichii, 9-18 parts of radix puerariae, 9-18 parts of salvia miltiorrhiza, 9-18 parts of astragalus membranaceus, 30-60 parts of leonurus, 9-18 parts of Chinese yam, 5-10 parts of radix paeoniae and 5-10 parts of rhizoma atractylodis. The traditional Chinese medicine compound preparation is capable of effectively treating animals poisoned by being fed with locoweed; and the problem that the existing treating medicines have toxicity is solved.

The invention discloses an oxytropis ehrig verticillium FEL5-AS1 of a synthesized swainsonine which is preserved in China Center for Type Culture Collection on 8th, Oct of 2008, with the preservation number of CCTCC NO: M 2008144. The colony of the bacterial strain is round, which is white at first. With the extension of the culturing time, the colony gradually changes into straw yellow, tan and pitchy, and the center processes a little. Mycelium has transverse layers but has no mediastinum. Part of aged mycelia produces conidiophores which have branches. The conidiophores bend like knee bending, and spore formation scars are obvious. Conidia and conidiophores have transverse layers but have no mediastinum. The diaphragm of the conidia is thick, and the transverse layers consist 1-4. The spores are bar-shaped, similar to column. The size of the conidia is 20-90Mum multiplied by 5-10 Mum. The bacterial strain is obtained from leaves, stems and seeds of astragalus strictus by artificial separation and screening and has the function of synthesizing swainsonine (SW) which is main toxin of locoweed.

The invention discloses an oxytropis ehrig sporangium FEL6-AS2 for synthesizing SW, which is preserved in the China Center for Type Culture Collection on October 8, 2008, with the preservation serial number of CCTCC No: M 208145. The colony of the strain is circular and is white originally, the color is gradually changed into a grey-white color, a grey-black color and a black color along with the prolongation of the culture time, and the center protrudes. The mycelium has transverse septa and does not have longitudinal septa, part of aged hyphae puts forth conidium peduncles, and the conidium peduncles have branches. Conidia and the conidium peduncles have the transverse septa and do not have the longitudinal septa, the transverse septa of the conidia are thicker, the number of the transverse septa is 1 to 4, sporules are in bar shapes and are approximately cylindrical, parts of the sporules are circular, and the size of the conidia is 20 to 80mum*6 to 12mum; and the strain is obtained from the leaves, stems and seeds of astragalus root with straight stems by artificial separation and screening, and has the function of synthesizing the SW which is the main toxin of locoweed.

The invention discloses an oxytropis Ehrig cinerea FEL2-OG which is preserved in China type culture collection center on October 18, 2008, and the preservation code is CCTCC NO: M 208142. Bacterial colony of bacterial strain is in a circular shape, is white in the beginning, and gradually changes to primrose yellow, yellow brown and black brown, and the centre of the bacterial colony slightly protrudes. Mycelium has transverse septa but has no mediastinums. Conidiophores peduncles grow on part of the aging hypha h, and the conidiophores have braches. Conidiophores and the conidiophores peduncles have transverse septa but have no mediastinums, the transverse septa film of the conidiophores is thicker, and the number of the transverse septa is 1 to 3; spores are in long rod shapes and are similar to column shapes, and the size of the conidiophores is 20 to 70 microns*5 to 10 microns. The bacterial strain is obtained from leaves, stems and seeds of oxytropis glacialis through manual separation and filtration, and has the function of synthesizing main toxin, that is swainsonine (Swainsonine, SW) of locoweed.

The invention discloses a pair of specific molecular detection primers for undifilum oxytropis and a method for detecting whether locoweed is infected by undifilum oxytropis with the pair of specific molecular detection primers for undifilum oxytropis. The pair of specific molecular detection primers include a base sequence equipped upstream primer OmtssuF and a downstream primer OmtssuR, the upstream primer OmtssuF and the downstream primer OmtssuR are 164bp products specifically amplified from undifilum oxytropis infected locoweed. According to the mtSSU coding sequence of undifilum oxytropis, the pair of specific molecular detection primers are designed. The detection primers have strong specificity and good sensitivity. The detection method provided by the invention carries out DNA amplification on the to-be-detected locoweed to obtain a 164bp strip, cannot amplify any product from other fungal DNA and locoweed DNA itself, can be used for fast, accurately and conveniently detecting whether a locoweed sample is infected by undifilum oxytropis, has strong specificity and good practicability, and has important significance to locoweed control.

The invention relates to a compound detoxifying agent for treating locoweed poisoning of livestock and a preparation method of the agent. The compound detoxifying agent is prepared by dissolving an antidote into 1000 parts of sterilization water, and the antidote is prepared from the following components in part by weight: 0.25-0.5 part of yeast mannan, 0.8-1 part of L-rhamnose, 3-5 parts of magnesium sulfate and 8-10 parts of sodium orthophosphate. The compound detoxifying agent for treating locoweed poisoning of the livestock with the formula provided by the invention has definite components, is proven to have no toxic and side effect of teratogenesis, mutagenesis, carcinogenesis and the like by safety toxicology experiments, and is safe and reliable when being applied to production.

The invention discloses a swainsonin antigen. The swainsonin antigen has a following chemical structural formula shown in the description, wherein BSA is bovine serum albumin. The preparation method of the swainsonin antigen comprises the steps that (1) swainsonin and ethyl malonyl chloride are subjected to a reaction to obtain a compound I shown in the description, the compound I is dechlorinatedunder the alkali function and acidified to obtain a compound II shown in the description, and the compound II and BSA are condensed to obtain the swainsonin antigen under the function of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1-hydroxybenzotriazole. The synthesis process of the swainsonin antigen is simple, easy to operate, low in cost and high in coupling ratio, the antigen can produce high-titer antibodies, and a new way is opened for the reasonable development and utilization of locoweed plants.

The invention relates to a block for preventing and treating locoweed poisoning of animals, belonging to the technical field of an animal medicine. The block consists of the following raw materials in parts by weight: 2-3 parts of molasses, 10-15 parts of common salt, 3-4 parts of quick lime, 6-7 parts of bran, 2-3 parts of copper sulfate, 10-20 parts of a trace element mixing agent, 5-10 parts of urea phosphate and 2-4 parts of a yeast mannan-phosphate-Mg2+ induction mixing agent, wherein the yeast mannan-phosphate-Mg2+ induction mixing agent consists of the following components by the following proportioning: yeast mannan, L-rhamnose, pharmaceutical-grade magnesium sulfate and pharmaceutical-grade sodium orthophosphate in weight ratio of (0.25-0.5): (0.8-1): (3-5): (8-10). The nutrition block is remarkable in treating effect, sufficient in various components and raw materials, good in safety, convenient to implement, reasonable in proportion and complete in nutrition.

The invention relates to a method for detecting a locoweed extractive and a content of swainsonine in a capsule. The method comprises the following steps: 1) extracting the locoweed extractive or swainsonine in the capsule via soaking through a formic acid-methanol solution with a volume ratio of 1%; 2) purifying the extracting solution by adopting a graphitized carbon solid phase extraction column and a syringe filter; 3) separating and detecting swainsonine via adopting a high performance liquid chromatogram-mass spectrometer (HPLC-MS); and 4) plotting swainsonine with different concentrations and corresponding peak areas to obtain a standard curve of swainsonine. The content of swainsonine is calculated by contrasting the peak area of a to-be-detected sample with the standard curve. The HPLC-MS is extremely high in sensitivity, is free of most matrix interference, and is more convenient and efficient in pre-treatment process, and accurate and reliable in detection result, compared with other detection methods. The method is high in sensitivity and good in specificity and reproducibility, and is a reliable analytical method.

The invention relates to the field of chemical treatment of natural medicines, in particular to a method for extracting swainsonine from barbadosweed endophytic fungi (U.oxytropis), which comprises the following steps: (1) preparing the barbadosweed endophytic fungi into a bacterial suspension, and culturing; (2) after culture of the bacterial suspension in the step (1) is finished, performing suction filtration, rinsing, drying, grinding and wall breaking, and collecting dry powder; (3) weighing the dry powder collected in the step (2), transferring the dry powder into a centrifuge tube, adding methanol, extracting, centrifuging, collecting supernate, evaporating to dryness, redissolving, filtering and diluting; according to the extraction method, the yield of swainsonine reaches 220 mu g/g and is remarkably improved compared with the prior art, the method is simple in process condition, safe, low in cost and easy for industrial production, a basis is provided for development and utilization of swainsonine produced through microbial fermentation, raw material resources of swainsonine products are expanded, and the method is suitable for industrial production. Wide application prospects are realized.

The invention relates to a coding gene of swainsonine degrading enzyme and application thereof, belonging to the fields of biotechnology and enzyme gene engineering. The coding gene of swainsonine degrading enzyme (gene AAur_2040) has a nucleotide sequence as shown in SEQ ID No. 1. A protein coded by the coding gene has an amino acid sequence as shown in SEQ ID No. 2. According to invention, the gene segment-- AAur_2040 responsible for degradation of swainsonine is cloned from Arthrobacter sp. HW 08 capable of effectively degrading swainsonine. The coding gene of swainsonine degrading enzyme and application thereof can provide effective biotechnological means for degradation of swainsonine and are of important significance to restoration of poisoning of animals caused by intake of locoweed.