95 results about "Hemolymph" patented technology

The present invention belongs to the field of feed additive medicine, in particular to the field of Chinese traditional herbal medicine used for the breeding of the South America white shrimp. The Chinese traditional herbal medicine used for the breeding of the South America white shrimp mainly comprises the components which is also combined in the weight ratio that: 10 to 15 of Chinese teasel root, 15 to 25 of largehead atractylodesrhizome, 10 to 18 of root of herbaceous peony, 10 to 15 of glossy privet fruit, 10 to 15 of rhubarb, 8 to 15 of honeysuckle flower, 8 to 15 of astragalus root and 3 to 10 of bupleurum. The present invention can increase the content of the lysozyme and the phagocytic power of the hemolymph, increase the levels of the SOD and the CAT, avoid the defects that the effect of the antibiotics alike medicines is not comprehensive and has certain side effects if used in amount, thus the 100 percent of the effective rate for preventing is reached, 73 percent of the highest survival rate of the South America white shrimp after using the medicine is realized.

The invention discloses an artificial medium of trichogramma. The artificial medium comprises the following ingredients in mass fraction: 19-40% of insect pupae hemolymph, 17-38% of egg yolk, 10-33% of milk, 4-19% of a Nissl salt solution, 0.8-8% of trehalose, 2-15% of a ganoderan solution with a mass fraction of 5% and 2-15% of a chitosan oligosaccharide solution with a mass fraction of 3.6%. By adopting the artificial medium of trichogramma, the breeding effect of trichogramma is obviously better than that of a traditional artificial medium through the trehalose, the ganoderan solution and the chitosan oligosaccharide solution via effective composition, and the content of insects can be reduced, so that the breeding cost of trichogramma is reduced. Through the artificial medium of trichogramma, trichogramma can be bred by subculturing, no degradation phenomenon exists, and the breeding effect is stable. By adopting the artificial medium of trichogramma, various kinds of trichogramma such as trichogramma dendrolimi, trichogramma confusum, Asian corn borer trichogramma, European corn borer trichogramma and geographical populations thereof can be bred, and good effects are realized.

The invention relates to the field of biotechnology, and particularly relates to the bioactivity of the insect, insect hemolymph and assembly active substance thereof in activating a prophenoloxidase activation system or / and Toll signal transmission path system and an application of the insect, insect hemolymph and assembly active substance thereof in microorganism and related molecular mode detection thereof, preparation of the assembly active substance of prophenoloxidase activation system and separation and purification of antibacterial peptide. The invention also provides an insect hemolymph collection method and the storage method and conditions of the hemolymph; under the conditions, the stability of the insect hemolymph is better, and the functions of the insect hemolymph can be better played.

Each 100 ml of blood anticoagulant contains trisodium citrate 3.31-3.32g, citric acid 0.01-0.05g, glucose 2.45-3.37g and the rest is water. The tests show that after the external blood is undergone the process of anticoagulation treatment it can prolong the stroage life of lymphocyte to 48-72 hr. from original 4-6 hr. Said invention also provides its application method.

The invention discloses application of glycine or a glycine metabolism regulator, i.e., 1-aminocyclopropane-1-carboxylic acid (ACPC), in improving domestic silkworm silk output, as well as a method for improving the domestic silkworm silk output by supplying exogenous glycine or ACPC to domestic silkworms in a larval period. According to the invention, by researching change in a hemolymph metabolite spectrum of different varieties of domestic silkworms having a large difference in silk output based on GC-MS metabonomics technology, strong correlation is proved to exist between the glycine (42.8% of silk fibroin), as an important component, of domestic silkworm silk fibroin, and a cocoon layer ratio (correlation coefficient is -0.892); the domestic silkworm silk output is significantly improved by intervening metabolism of the silkworms in the larval period through in vitro administration of the glycine or the glycine ACPC; therefore, the invention not only widens application of the glycine or the ACPC in the field of biology, but also provides a new method for improving the silk output in silkworm industry.

The invention discloses a pteromalus puparum venom Kazal-type serine protease Inhibitor PpSPI20 protein. The protein comprises an amino acid sequence shown as SEQ ID NO: 2. The invention further discloses genes coding the pteromalus puparum venom Kazal-type serine protease Inhibitor PpSPI20 protein. The genes comprise a nucleotide sequence at sites 70-246 in SEQ ID NO:1. The inhibitor disclosed bythe invention can be used for inhibiting activation of hemolymph PPO (Poly Phenylene Oxide) of imported cabbageworms or papilio xuthus. The invention further discloses a method for improving prevention ability of brassicaceous vegetables on lepidoptera pests. The method comprises the following steps: transforming brassicaceous vegetable cells by using the genes comprising the nucleotide sequenceshown as SEQ ID NO:1, and cultivating the transformed brassicaceous vegetable cells into plants.

The invention discloses a method for determining the contents of albendazole and metabolites thereof in hemolymph of Bombyx mori by using UFLC-MS / MS. The method comprises the following steps: (1) extracting hemolymph of Bombyx mori so as to obtain a to-be-measured sample solution; (2) measuring standard analytical data of standard albendazole and metabolites thereof; (3) injecting the to-be-measured sample solution obtained in the step (1) into an ultra-fast liquid chromatograph for separation and analysis according to a method used in the step (2) and detecting substance signals by using a triple-quadrupole tandem mass spectrometer so as to obtain analytical data of a to-be-measured sample; and (4) comparing the analytical data of the to-be-measured sample obtained in the step (3) with chromatographic analysis data of albendazole and metabolites thereof so as to realize fast qualitative and quantitative determination of albendazole and metabolites thereof in hemolymph of Bombyx mori.

The invention discloses a chemical derivation-based detection method for hemolymph metabolin of migratory locust, and relates to the field of animal ecology and analytical chemistry. The detection method specifically comprises the following steps of: taking hemolymph, deproteinizing by adding ethanol-acetonitrile mixed solvent, supersonically extracting, and centrifuging at a high speed to obtain metabolin extracting solution; drying the extracting solution by means of pressure reduction, adding methoxamine hydrochloride-pyridine solution, supersonically dissolving, blending by means of vortex, and deriving in an oximation way in a constant-temperature water bath; adding N-methyl-N-trimethylsilane-trifluoroacetamide reagent, blending by means of vortex, and deriving in an alkylation way in the constant-temperature water bath; and adding methyl oleate-normal heptane solution to compensate the volume, blending by means of vortex, and analyzing by a spare gas chromatography-mass spectra coupling technique. The detection method provided by the invention is convenient, high-efficiency, mild in reaction conditions, good in repeatability, wide in metabolin coverage, suitable for developing the multi-center and large-sample research, and capable of providing help for knowing the biological law of various migratory locust variations and the occurrence mechanism of the locust plague.

The invention provides barley worm natural antibacterial protein and a preparation method and application thereof. According to the barley worm natural antibacterial protein and the preparation method and application thereof disclosed by the invention, escherichia coli and / or staphylococcus aureus are / is used as an inducer, after bacterial liquid is injected to barley worm larvae or yellow mealworm larvae, then the barley worm larvae or the yellow mealworm larvae are raised for some time, hemolymph of the larvae is collected, and antibacterial protein is extracted. The obtained antibacterial protein can be used for preparing an antibacterial agent or a preservative. The antibacterial protein provided by the invention has the characteristics of being good in broad-spectrum antibacterial properties, high in stability, good in safety and the like, the preparation technology is simple, raw materials are cheap and easy to obtain, and the application prospect is great.

The invention relates to a shellfish hemolymph cell classification method based on multiple technologies. The shellfish hemolymph cell classification method comprises following steps: hemolymph collection and total haemocyte counting, cell optical microscope analysis, flow cytometry, transmission electron microscopy analysis, scanning electron microscope analysis, and statistic analysis. The shellfish hemolymph cell classification method is provided based on the prior art; deficiencies are avoided; problems, that complicating of shellfish hemolymph cell classification is caused by the interspecific difference of shellfish and the difference of research methods and unified standards are difficult to achieved, are avoided; and the shellfish hemolymph cell classification method possesses important meaning on shellfish hemolymph cell classification and study on the action of shellfish hemolymph cells in body immunity process.

The invention relates to the technical field of biological prevention and control, in particular to a novel method for preventing and controlling brown planthopper. Trifloxystrobin-tebuconazole water dispersible granule (the mass percentage of effective components is 75 percent, wherein the mass percentage of trifloxystrobin is 25 percent, and the mass percentage of tebuconazole is 50 percent) is used for preventing and controlling brown planthopper filial generations. The most remarkable characteristic of the novel method is that yeast-like symbiotes are prevented or inhibited from entering ovarian tissues from the hemolymph of brown planthopper female adults through a trifloxystrobin-tebuconazole sterilizing agent, so that the reproductive systems of the brown planthopper is influenced, the egg laying amount is reduced, and the hatching rate of the laid eggs is remarkably lowered due to the lack of symbiotic bacteria. Thus, the generation of brown planthopper filial generations is effectively controlled.

The invention provides an artificial intelligence identification method for a peripheral hemolymph micronucleus cell image, and the method comprises the steps: collecting a to-be-analyzed sample through a collection device, and obtaining a cell image; carrying out segmentation processing on the obtained cell image to obtain a foreground to-be-detected area; identifying, classifying and counting the cells on the foreground to-be-detected area; storing the cell type information on the foreground to-be-detected area and the position information of the foreground to-be-detected area on the cell image to form traceable information; and forming an image-text report according to the traceable information, and storing the image-text report as an artificial intelligence system learning training material. According to the artificial intelligence identification method for the peripheral hemolymph micronucleus cell image provided by the invention, automatic identification and classification of specific images of lymphoblasts, micronucleus cells and nude nuclei of peripheral blood can be realized. Compared with an existing microscope artificial microscopic examination method, the statistical accuracy is higher, and the detection speed is remarkably increased.

The invention discloses a method for estimating the toxicity contribution rate of metal nanoparticle ions and a nano effect and belongs to the field of artificial metal nanoparticle toxicity effect evaluation. Hemolymph cells of shellfish in an ocean are selected as an experimental subject, cell culture mediums containing metal ions or nanoparticles are set as two experimental groups according to the content of metal ions released by the metal nanoparticles, and the toxicity contribution rates of an ion effect and the nano effect to the metal ions and the nanoparticles are researched. The nano effect and the ion effect on the metal nanoparticles are researched for the first time, the hemolymph cells of the immune system of a benthic organism in the ocean are selected as the experimental subject, and the cells are very sensitive to stress of pollutants; besides, multiple physiological indexes are selected, the research level is pushed to the molecular immune level, and the effectiveness and the accuracy of toxicity effect evaluation on artificial metal nanoparticles are improved.

The invention relates to a preparation method of silkworm decellularization hemolymph liquid. The preparation method comprises a series of steps of disinfecting the body surfaces of silkworms, preparing the hemolymph liquid of the silkworms, carrying out antioxidant treatment and cell extraction on the liquid, removing bacteria from the liquid and the like. The method takes the hemolymph liquid of the silkworms as materials to prepare a nutrient substance applied to the cell culture of insects. Vitamin C is used for inhibiting the activity of tyrosine oxidase in the hemolymph liquid, so that the hemolymph liquid does not get black by oxidation when being contacted with the outside, the composition and the content of main nutrient substances in the hemolymph liquid are not changed, and the cell culture is not influenced. Compared with serum, the hemolymph liquid of the silkworms is more suitable for the cell culture of the insects, and is low in price.

The invention discloses a Pteromalus puparum venom Kazal serine proteinase inhibitor polypeptide which has an amino acid sequence represented by SEQ ID NO:2. The invention simultaneously discloses gene coding the Pteromalus puparum venom Kazal serine proteinase inhibitor polypeptide, wherein the gene has a nucleotide sequence represented by a 145-297 nucleotide sequence in SEQ ID NO:1, or has a nucleotide sequence having at least 70% homology with the 145-297 nucleotide sequence in the SEQ ID NO:1, or has a nucleotide sequence which can be hybridized with the 145-297 nucleotide sequence in the SEQ ID NO:1 at 40-55DEG C. The polypeptide of the invention is used for preparing Pteromalus puparum venom Kazal serine proteinase inhibitors, and the inhibitors can be used for inhibiting the activation of Pieris rapae or Papilio xuthus hemolymph PPO (prophenoloxidase).

The present invention relates to a preparation method of a snail respiratory protein. A respiratory protein containing copper ions is extracted from a snail vivo used as an intermediate host of Schistosoma japonicum by virtue of mechanical extrusion, tissue dissection, hemolymph extraction, ammonium sulfate precipitation, super high-speed centrifugal separation and other means. The respiratory protein contains 0.2-0.3% of copper, the UV characteristic absorption peak is at 275nm and 340nm, and a cylindrical agreegate structure is observed under a transmission electron microscope.

The invention discloses a method for constructing transgenic Nosema bombycis, characterized by injecting a transgenic vector wrapped by liposome in body cavity of the Bombyx mori infected with Nosema bombycis, after 2-4 days, adding silkworm hemolymph in Bombyx mori BmN culture cells to cultivate; when microsporidian presents green fluorescent light, using an insect medium to extremely dilute and adding into a cell culture plate, selecting a culture pore where only one culture cell presents green fluorescent light, adding normal culture cells to culture for 2-4 days to obtain infected culture cells; repeating for at least three times, taking the obtained infected culture cells, conducting homogenate and crushing the cells, conducting differential centrifugation, purifying microsporidian, infecting the Bombyx mori, after the Bombyx mori falls ill, purifying the infected tissues to obtain the transgenic Nosema bombycis. According to the invention, exogenous genes can be integrated into the Nosema bombycis genome, the modification of Nosema bombycis by gene engineering is realized, the adjustment of expression of the microsporidian genes is realized, and a method for microsporidian gene function research is provided.

The invention relates to the technical field of biological prevention and control and especially relates to a use of a fungicide in prevention and control of brown planthopper filial generation. The fungicide which has toyocamycin, tetramycin P, tetrin B and tetramycin A total content of 27% is used for prevention and control of brown planthopper filial generation, wherein toyocamycin content is 5.58% by mass, tetramycin P content is 7.09% by mass, tetrin B content is 4.44% by mass and tetramycin A content is 9.64% by mass. The fungicide comprising toyocamycin, tetramycin P, tetrin B and tetramycin A can prevent or inhibit yeast-like symbiote entrance into ovarian tissue from female brown planthopper adult hemolymph thereby influencing a brown planthopper reproductive system, reducing egg laying amount and substantially reducing egg hatchability because of symbiotic bacterium loss so that brown planthopper filial generation is effectively controlled.

The invention belongs to the technical field of plant poison toxicity evaluation, and provides a metabonomics-based research method for a toxicity effect of jujube flower nectar on honeybees, aiming at the current situation that the toxicity effect of the jujube flower nectar on the honeybees cannot be explained in the existing research. The research method comprise the steps of: collecting honeybees with jujube flower diseases and taking healthy honeybees as a contrast; collecting hemolymph of the two groups of honeybees; combining a UPLC-Q-TOF MS technology with multi-dimensional statisticalanalysis to carry out full spectrum analysis on the sample to obtain primary mass spectrum data and secondary mass spectrum data; adopting xCMS software to carry out peak extraction and metabolite identification on the data, screening metabolite targets and metabolic profiles related to honeybee jujube flower diseases from metabolite, tracking and screening of pathogenicity of jujube flower nectar on honeybees under the guidance of metabolite target analysis and metabolic profile analysis, and illustrating a toxic action mechanism of the jujube flower nectar on honeybees. The research methodhas the advantages of being low in cost, easy to operate, rapid in detection, high in sensitivity, strong in selectivity, good in repeatability, comprehensive in obtained information and the like.

The invention provides a simple and high-efficiency method for inducing spiders to produce antibiotic active substances, which comprises the following steps: a. ultraviolet irradiation is used for inducement: larinioides cornuta is put into a culture vessel and irradiated for 0.5-3 hours under 20W ultraviolet light and in the distance of 1 meter; then the larinioides cornuta is raised continuously; b. hemolymph of the spiders 1-120 hours after induced is extracted and collected by applying a tissue homogenate method; c. B bronchiseptica is employed as indicator bacteria in a coated flat drilling method to test antibacterial activity of the hemolymph. The antibacterial activity of extracted spider hemolymph after irradiated and induced by ultraviolet is high, and the antibiotic active substances can be applied to development of antibiotics.

The invention belongs to the technical field of freshwater aquaculture, and relates to a method for analyzing aeromonas hydrophila infected macrobrachium nipponense hemocyte differential expression protein based on a proteomics quantitative technology. The method comprises the following steps of (1) extracting the hemolymph of macrobrachium nipponense infected with aeromonas hydrophila, separatingthe blood cells, and cracking the cells to extract protein; (2) carrying out trypsin enzymolysis on the protein obtained in the step (2), and carrying out iTRAQ labeling on an enzymolysis peptide fragment; (3) carrying out purification, liquid chromatographic separation and mass spectrometric analysis on the peptide fragment marked in the step (3); and (4) carrying out protein identification, quantitative analysis and bioinformatics analysis on the mass spectrum analysis result obtained in the step (3), and analyzing to obtain differentially expressed protein. The invention is based on the rapid proteomics research of the infected macrobrachium nipponense by iTRAQ, and provides a reference for further researching the effect of immune-related proteins.

The invention discloses a pteromalus puparum venom Kazal-type serine protease inhibitor protein PpSPI24 with an amino acid sequence as shown in SEQ ID NO:2. The invention further discloses a gene forcoding the chrysalis nasonia venom Kazal-type serine protease inhibitor protein PpSPI24, and the gene has nucleotide sequences of sites 70-231 in SEQ ID NO:1. The inhibitor can be used for inhibitingactivation of imported cabbageworm or papilio xuthus linnaeus hemolymph PPO. The invention further discloses a method of improving prevention force, on lepidoptera pests, of brassicaceous vegetables,and the method comprises the following steps: converting cells of the brassicaceous vegetables by the gene with the nucleotide sequences as shown in SEQ ID NO:1; and cultivating the converted cells ofthe brassicaceous vegetables into plants.

The invention discloses a shelling treatment method for tegillarca granosa. The method is characterized in that the method includes the following steps: placing tegillarca granosa into a polyethylenebag holding an aqueous solution; placing the polyethylene bag into an ultra-high pressure setting, control pressure being 300-400 MPa; and performing pressure maintaining for 1-5 min so that the shelling treatment of the tegillarca granosa can be accomplished, wherein the aqueous solution is the aqueous solution containing 0.30-0.60 wt% of ascorbic acid. The advantages of the method are as follows: effective sterilization can be realized; the shelling efficiency of the tegillarca granosa can be enhanced; the complete separating of meat can be realized; hemolymph liquid can be completely reserved in tissues; and the original flavor and nutrient compositions of the tegillarca granosa meat can be maintained.

A method for simply and rapidly detecting oyster ploidy is disclosed. The method includes firstly crumbing shell edges, of two shells of an oyster, close to the adductor muscle with sharp-nose pliersto form a gap having a length of 2-3 mm; drawing hemolymph by inserting a disposable syringe through the gap into a hemolymph sinus of the adductor muscle; after the hemolymph is immobilized with absolute alcohol, performing centrifugation and removing a liquid supernatant; adding PBS and suspending the mixture again; performing RNA enzyme reaction and PI staining in order; and performing filtration with a cloth sieve having a size of 300 meshes; and then detecting the DNA content with a flow cytometer to determine the ploidy. The method performing ploidy detection through drawing the hemolymph in the hemolymph sinus of the adductor muscle of the oyster is non-lethal, simple, rapid, not limited by oyster living environment, and the like, and is of great significance for oyster polyploidy breeding research, especially ploidy detection of live oyster polyploidy and subsequent deep research.

The invention discloses a preparation method of an apis cerana antimicrobial peptide AccRoyalisin polyclonal antibody and the use thereof. Apis cerana royal jelly antimicrobial peptide AccRoyalisin recombined and expressed by escherichia coli is taken as an antigen, and a big rabbit is immunized to prepare a polyclonal antibody, the valence of the antibody is larger than 40,000 times, the polyclonal antibody is taken as a primary antibody, recombined and expressed GST-AccRoyalisin is taken as a standard protein antibody to perform doubling dilution, the absorbance of GST-AccRoyalisin protein samples with different concentrations is detected through an Elisa method by a microplate reader, and the standard curve between the absorbance and the MRJPI protein concentration is established. Different bee species or different bee swarms of hemolymph and lyophilized royal jelly powder serve as antibodies after being diluted, and the absorbance of the obtained sample is detected through adopting the Elisa method, and is introduced into the established standard curve to calculate the content of Royalisin of the obtained sample. The invention provides the new use for the Royalisin antibody, and provides a novel quick, qualitative and quantitative detection method for evaluating premunition of the bee species and bee swarms in the bee-keeping field, breeding the antiviral bee swarms and evaluating the antiseptic and antibiosis performance of royal jelly.

The invention relates to an efficient biomolecular efficient delivery technology, and aims to provide a nilaparvata lugens vitellogenin N-terminal polypeptide (VgNPP) with an ovary targeting functionand application of the nilaparvata lugens VgNPP. An amino acid sequence of the polypeptide is shown in SEQ ID NO: 1. According to the application of the polypeptide as a nilaparvata lugens ovary-targeting biomolecule transporter, the nilaparvata lugens VgNPP carries a biomolecule to enter an egg through nilaparvata lugens parent hemolymph in a targeting manner; and the biomolecule is any one of ora complex of more of nucleic acid, oligopeptides, polypeptides, protein, saccharides, lipid, nanoparticles and a small-molecule compound. The popypeptide provided by the invention is used as the biomolecule transporter, particularly as a gene editing element transporter to carry the biomolecule to enter the ovary of the nilaparvata lugens. Compared with other foreign high polymer materials or foreign protein, the VgNPP is derived from the nilaparvata lugens, so that the VgNPP has no toxic or side effects on the nilaparvata lugens and has very broad application prospects.

The invention relates to the technical field of biological control, in particular to application of nativo to control over filial generations of brown planthopper. Native water dispersible granule (with 75 % mass percent of active ingredients including 25 % mass percent of trifloxystrobin and 50% mass percent of tebuconazole) is utilized by the invention for control over filial generations of brown planthopper.The most salient feature of the invention is described in that nativo, as bactericide, helps to prevent of inhibit yeast-like endosymbiote from entering ovarian tissues from hemolymph of female adults of brown planthopper such that a genital system of brown planthopper is affected and the egg laying amount is reduced; and the hatching rate is greatly decreased because produced eggs lack symbiotic bacteria so that generation of the filial generations of brown planthopper is effectively controlled.

The invention discloses a method for a direct polymerase chain reaction (PCR) adopting litopenaeus vannamei hemolymph. The method comprises the steps of 1, anticoagulant preparation; 2, hemolymph collection and pretreatment; 3, PCR treatment. The method has the advantages of being easy to operate, low in cost, safe and free of toxins, the complex step of extracting DNA is omitted, harm to parent prawns is avoided, and the modern biological requirements are met.

The present invention is directed to methods for collecting a commercially useful quantity of crude hemocyanin from live gastropod molluscs by isolating hemolymph in a sinus of the animal followed by extraction of the hemolymph. The methods of the invention do not require incision of the vascular system or injury or death to the animal. In addition, the methods of the invention enable the periodic extraction of hemolymph from the same source animals.