210 results about "White spot syndrome" patented technology

The invention discloses an antiviral complex traditional Chinese medicine and a preparation method and application of the antiviral complex traditional Chinese medicine. The complex traditional Chinese medicine comprises isatis root, rheum officinale, astragalus mongholicus, folium isatidis, houttuynia cordata and polygonum cuspidatum. The antiviral complex traditional Chinese medicine and the preparation method provided by the invention are reasonable in prescription, and simple in preparation technology; as the clinical test shown, the traditional complex traditional Chinese medicine can be applied to preparing medicine for treating or preventing white spot syndrome of cray, hemorragic disease of grass carp, haematopoietic necrosis of crucian, koi herpes virus disease, channel catfish virus disease, iridovirus disease of siniperca chuatsi, and virosis of aquatic livestock; and the antiviral complex traditional Chinese medicine has high efficiency, is safe and has no toxic and side effect, and has a wide clinical application prospect.

The invention belongs to the technical field of marine organism pathogen detection, and in particular relates to a multiple-PCR detection method and a kit for simultaneously detecting WSSV (white spot syndrome virus), AHPND (acute hepatopancreatic necrosis disease), EHP (enterocytozoon hepatopenaei) and IHHNV (infectious hypodermal and hepatopancreatic necrosis virus) in prawns. With the application of the multiple-PCR detection method, amplification of four pathogen DNAs can be simultaneously conducted in one time by virtue of special detection primers, namely WSSV F and WSSV R, AHPND F and AHPND R, EHP F and EHP R as well as IHHNV F and IHHNV R of the WSSV, the AHPND, the EHP and the IHHNV. The multiple-PCR detection method provided by the invention is convenient and rapid, low in detection limit, strong in specificity, high in sensitivity and high in accuracy rate and is capable of simultaneously detecting the white spot syndrome virus, the acute hepatopancreatic necrosis disease, the enterocytozoon hepatopenaei and the infectious hypodermal and hepatopancreatic necrosis virus in the prawns.

The invention discloses a Chinese herbal medicine immunopotentiator for resisting white spot syndrome virus (WSSV) of procambarus clarkia, as well as a preparation method and an application thereof. The preparation method comprises the following steps: milling rheum officinale, epimedium, astragalus membranaceus and isatis root and fully mixing the ingredients in proportion to prepare the powdery immunopotentiator. The Chinese herbal medicine immunopotentiator disclosed by the invention is stable and wide in raw material resources, simple to prepare, and low in cost, is based on the immunomechanism of shellfish to provide Chinese herbal medicines for the shellfish, and can be used for improving the immunity and oxidation resistance of a body during application, improving the capacity of the procambarus clarkia for resisting WSSV infection, and promoting the growth of the procambarus clarkia.

An expression cassette, wherein said expression cassette comprises: (a) an ie 1 promoter from white spot syndrome virus or a variant thereof; (b) a nucleic acid sequence encoding an N-terminal gp64 signal peptide or variant thereof, a nucleic acid sequence encoding an antigenic peptide, a nucleic acid encoding a transmembrane region, and a nucleic acid encoding a gp64 cytoplasmic region or variant thereof; in which the promoter is operably linked to the nucleic acid sequence.

A compound Chinese herbal medicament preparation for preventing and treating white spot syndrome is prepared by following raw materials by weight: 20-35 parts of Corydalis bungeana Turcz., 25-40 partsof radix isatidis, 15-40 parts of radix astragali, 20-40 parts of dyers woad leaf, 25-35 parts of wrinkled giant hyssop, 10-30 parts of Japanese eupatorium, 15-25 parts of roughhaired holly root and5-15 parts of Gynostemma pentaphylla. The compound Chinese herbal medicament preparation can effectively prevent and treat the white spot syndrome. The invention also discloses a forage comprising thecompound Chinese herbal medicament preparation and a method for feeding the same. The forage comprising the compound Chinese herbal medicament has simple processing technology and convenient feeding.The method for feeding the forage comprising the compound Chinese herbal medicament is scientific and reasonable and has a good antiviral effect.

The present invention is prawn white spot syndrome virus gene diagnosing kit and its usage. The kit and its usage is designed based on two pairs of primers for the conservation sequence of prawn white spot syndrome virus gene. By means of PCR technology, the present invention detects qualitatively the specific DNA nucleic acid segment of prawn white spot syndrome virus fast, simply, specifically and sensitively. The present invention may be used in the tracing detection of different stages of prawn cultivation and environment monitoring.

The present invention relates to a composition useful as prophylactic and / or therapeutic agent for the management of viral and bacterial diseases in aquatic animals, said composition containing effective amount of extract obtained from the plants Lantena camera, Aegle marmelos, Ocimum sanctum, Mimosa pudica, Cynodon dactylon, Curcuma longa, and Allium sativum, optionally in combination with a pharmaceutically acceptable carrier, diluents or excipients.

The invention relates to a method for preparing a recombining spore of surface display foreign protein, in particular to a recombining spore for surface display prawn white spot syndrome virus Vp28 protein of a bacillus subtilis spore. In the method, a bacillus subtilis spore capsid protein gene is used as a molecule vector and is recombined with an antigen protein gene on the surface of the prawn white spot syndrome virus to construct an integral recombining plasmid in fusion expression; the bacillus subtilis is converted to a bacillus subtilis recombining strain; and the recombining spore of the spore surface display prawn white spot syndrome virus protein produced by the recombining strain is induced.

The invention relates to a novel multivalent carrier vaccine for a shrimp. The vaccine is characterized by comprising a bacillus subtilis recombination strain obtained by a genetic engineering technique, the strain belongs to the category of bacillus subtilis HT5303 and is preserved in China Center for Type Culture Collection, and the preservation number is CCTCC NO.: M 2012395. When spores are formed by the bacillus subtilis vaccine strain, white spot syndrome virus VP281 protein can be demonstrated on the surfaces of the spores, and fusion protein of cell-penetrating peptides, white spot syndrome virus VP19 and vibrio harveyi 3-glyceraldehyde-phosphate dehydrogenase (GAPDH) can be secreted and expressed when vegetative cells are formed. Compared with other vaccines, the multivalent carrier vaccine has the advantages of cross protection and long protection period. Moreover, the vaccine can be added into feeds and has the advantages of resistance against high-temperature setting of feeds and broad prospects for commercial development.

The invention discloses an engineered protein TAT-VP28 against white spot syndrome virus of shrimp and its preparation and purpose. The sequences of the engineered protein TAT-VP28 provided by the invention are nucleotide sequence as shown in SEQIDNO:1 and amino acid sequence as shown in SEQIDNO:2, and the bacterial strain is a recombinant gene engineering bacterial strain Escherichiacoli BL21(Pet-28a-TAT-VP28). WSSV is extracted from shrimp bodies and the VP28 gene is obtained through PCR; TAT-VP28 sequences are amplified by the PCR method and then the recombinant plasmid pET-28a(+)-TAT-VP28is constructed, followed by the transformation of the recombinant plasmid pET-28a(+)-TAT-VP28 into the Escherichia coli BL21(DE3). The gene engineering fusion protein can be applied to preparing medicaments for treating or preventing the white spot syndrome virus of shrimp, has large expression amount and soluble performance, is easy for industrial production at low cost with good security, and has a better lasting protective effect of preventing the white spot syndrome of shrimp.

The invention discloses a genetic engineering protein TAT-VP28-GH capable of resisting white spot syndrome virus (WSSV) as well as preparation and application thereof. The sequences of the genetic engineering protein are a nucleotide sequence shown in SEQIDNO:1 and an amino acid sequence shown in SEQIDNO:2; and the strain of the genetic engineering protein is a recombinant genetic engineering strain EscherichiacoliBL21pEt-32A-TAT-VP28-GH. The preparation method comprises the following steps: extracting WSSV from prawn, and obtaining VP28 gene by a polymerase chain reaction (PCR); amplifying aTAT-VP28 sequence through the PCR; extracting total mRNA (messenger RNA) from hypophysis cerebri of carp, and obtaining total cDNA through RT-PCR (reverse transcription-PCR); obtaining a mature peptide gene of carp growth hormone through PCR, inserting acid hydrolysis site aspartate and proline (Asp-Pro-Asp) in the upstream of the mature peptide gene; then constructing a recombinant plasmid pEt-32A-TAT-VP28-GH; and transforming the recombinant plasmid pEt-32A-TAT-VP28-GH to Escherichia coil BL21, and efficiently expressing fusion protein TAT-VP28-GH in a dissoluble form under the induction of isopropyl beta-D-1-thiogalatopyranoside (IPTG). The fusion protein can effectively prevent the prawn from suffering WSSV and promoting growth development of the prawn.

The invention discloses a combined method of a stereo ecological polyculture mode and the prevention and control of hepatopancreatic necrosis disease and white spot syndrome. Specifically, 3 days before adding prawn seeds, 200 scatophagus argus fries with length of 3-5cm and 300 siganids fries with length of 2-3cm are added into every Mu of a breeding pond; when prawn seeds are to be added, a prawn seed addition amount is 80000-100000 seeds / Mu; when prawn length reaches 4-5cm, 5-6 Nile perch with length of 8-10cm and 10-15 hybrid giant tiger groupers with length of 8-10cm are added into every Mu of the pond. According to the invention, fishes with different feeding habits are cultured with prawns at a same time, such that diseases and weak prawns are eliminated in time, prawn disease infection source is eliminated, and a water cleaning function is realized. A polyculture mode of prawns and fishes with suitable specifications is started from a breeding early-mid stage, such that a prawn disease infection source is cut off, and hepatopancreatic necrosis disease and white spot syndrome outbreak is prevented. The breeding mode is optimized, such that prawn breeding success rate is improved. The method provided by the invention is suitable for the breeding of prawns such as litopenaeus vannamei, marsupenaeus japonicas, penaeus monodon and the like.

The present invention relates to a novel composition useful for inhibiting White Spot Syndrome Virus (WSSV) infection of crustacean animals, particularly those of the genera Penaeus sp. More specifically, the novel composition comprises a polypeptide whose amino acid sequence corresponds to at least a portion of Vp28, a surface protein of WSSV, or an antibody that specifically binds the polypeptide. The polynucleotide sequences encoding the Vp28 polypeptides of the present invention are also disclosed. Further disclosed are methods for using the novel compositions to inhibit WSSV infection in crustacean animals.

The invention relates to an anti-white spot syndrome virus (WSSV) autophagy associated gene Cq-Atg8, and a preparation method and application thereof. The anti-WSSV autophagy associated gene Cq-Atg8 is from C. quadricarinatus and is named Cq-Atg8. The preparation method comprises the steps of (1) constructing a Cq-Atg8 pronucleus recombinant expression vector; (2) leading the Cq-Atg8 pronucleus recombinant expression vector obtained in the step (1) into a host cell, and carrying out inducible expression on the host cell by using isopropyl-betad-thiogalactoside (IPTG), so as to obtain an expression product; and (3) separating and purifying the expression product obtained in the step (2) so as to obtain the recombinant protein (rCq-Atg8). The rCq-Atg8 can inhibit infection and duplication of WSSV remarkably, and therefore the rCq-Atg8 can be used in preparation of anti-WSSV medicines and anti-disease feed additives for animals.

The invention discloses a White Spot Syndrome Virus(WSSV) resisting engineering protein VP28-CD, the preparation and application thereof. The engineering protein VP28-CD is an isolated protein with sequence of ribonucleotide of SEQ ID NO:1 plus the amino acid sequence of SEQ ID NO:2; the bacterial strain is recombinant gene engineering scherichia coli BL21 (pET-32a-VP28-CD), CCTCC No:M208032; White Spot Syndrome Virus(WSSV) is isolated from prawn and gene VP28 is obtained through PCR; silkworm is induced with colon bacillus JM109 to obtain the total mRNA of fat body of the silkworm and the total cDNA is obtained through RT-PCR; mature gene of silkworm antibacterial peptide Cecropin D is obtained through PCR, with the upper reach inserted with aspartic acid-proline at the acid hydrolysis locus; after that recombinant plasmid Pet-32a(+)-VP28-CD is constructed; the recombinant plasmid Pet-32a(+)-VP28-CD is transformed into colon bacillus BL21(DE3); under IPTG induction, the fused protein VP28-CD achieves soluable and high-level expression. The fused protein is applicable in the preparation of the drugs for curing and preventing White Spot Syndrome Virus(WSSV).

The invention discloses a feed additive capable of reducing the white spot syndrome virus disease of penaeus vannamei boone. The feed additive comprises the following components in parts by weight: 0.5-2 parts of photosynthetic bacteria, 0.2-1.5 parts of bacillus subtilis, 1-3 parts of glucose, 2-4 parts of Chinese honey locust, 3-7 parts of purple perilla, 5-7 parts of green tangerine peel, 1-4 parts of cassia twig, 3-7 parts of white paeony roots, 0.01-0.5 part of vitamin C, 0.01-0.05 part of vitamin K and 15-20 parts of a carrier. 0.1-1 percent by weight of the feed additive disclosed by the invention is added into a conventional mixed feed for a prawn, and the obtained feed can be used for feeding the penaeus vannamei boone. By the adoption of the feed additive disclosed by the invention, the occurrence of the white spot syndrome virus disease in the cultivation process of the penaeus vannamei boone can be remarkably reduced, the cultivation survival rate is improved, the quality of a prawn body is improved, the water quality is improved, and the economical and ecological benefits are increased.

The invention discloses an anti-prawn white spot syndrome virus (WSSV) recombinant Salmonella typhimurium strain, and a preparation method and an application thereof. The preparation method comprises the steps of: extracting WSSV genomic DNA from an infected prawn, and obtaining WSSV envelope protein VP28 genes by a PCR method; extracting total RNA from a silkworm fat body, and obtaining silkworm antimicrobial peptide CD genes by a reverse transcription PCR method; constructing an eukaryotic expression plasmid pcDNA3.1(-)-VP28-CD containing the WSSV envelope protein VP28 genes and the silkworm antimicrobial peptide CD genes; and transforming into attenuated Salmonella typhimurium W0420 strain competent cells. The WSSV recombinant Salmonella typhimurium strain can be used for preparing a vaccine, which can be directly orally fed prawns. Compared with the current commonly used protein vaccines and injectable DNA vaccines, the inventive vaccine has double functions of inhibiting WSSV infection and enhancing prawn immunity.

The invention discloses a kit for detecting white spot syndrome virus (WSSV) of prawn. The kit comprises DNA extractant of a sample to be detected, LAMP pre-reaction liquid, Bst DNA polymerase, LAMP reaction stabilizing solution, indicator, positive control DNA and negative control DNA. The method also discloses a method for detecting WSSV by utilizing the kit. The kit of the invention has strong specificity and can be applied to scientific research and WSSV detection of cultivation site. The method of the invention is low in cost, and is rapid and sensitive, and the detection result can be judged directly by naked eyes.

The invention relates to a method for breeding the FanNaBing sterilize which can resist hickie complex symptom virus. Wherein, it comprises: (1) selecting parent shrimp while the female-male ratio is 1:2; (2) counteracting toxic substances that ejecting hickie complex symptom virus solution into the first section of muscle at the back of parent; (3) eliminating the ill and dead parents, to select the FanNaBing sterilize which can resist hickie complex symptom virus; (4) breeding parent shrimps.

The invention discloses a method for controlling a white spot syndrome by culturing hybrid giant tiger groupers and marsupenaeus japonicas in a mixed mode. The method comprises the steps that marsupenaeus japonicas seeds with the overall length of 0.8-1.0 cm are put into a pond, the putting quantity is 70,000-80,000 per mu, the hybrid giant tiger groupers with the body length of 8-10 cm are put into the pond when the marsupenaeus japonicas seeds are cultured until the body length of each marsupenaeus japonicas seed is 4-5 cm, and the putting quantity is 10-15 per mu. According to the method, according to the life habits of the hybrid giant tiger groupers and the marsupenaeus japonicas, by putting the hybrid giant tiger groupers in the appropriate density and appropriate specification into the high-density intensive culturing pond of the marsupenaeus japonicas, occurrence of the diseased and dead marsupenaeus japonicas can be reduced in the marsupenaeus japonicas culturing process, a good ecological culturing environment is supplied to growth of the marsupenaeus japonicas, the survival rate of the marsupenaeus japonicas is increased, the incidence is decreased, the culturing success rate is high, and the method is suitable for development of scale culturing of the marsupenaeus japonicas in China.

The invention discloses an envelope protein VP28 idiotype monoclonal antibody against shrimp white spot syndrome virus (WSSV) and a preparation method thereof. The antibody is secreted by a hybridoma cells with the collection number of CCTCC-CT200938, is prepared by taking anti-WSSV-VP28 monoclonal antibody (Ab1) as antigen, can bind with anti-WSSV-VP28 antibody of hare, and has the capability of competing with WSSV to bind with the anti-WSSV-VP28 antibody of the hare. The anti-WSSV-VP28 idiotype monoclonal antibody (Ab3) prepared by taking the antibody as antigen can bind with the WSSV, the binding site of the anti-WSSV-VP28 idiotype monoclonal antibody (Ab3) and the WSSV is located on an envelope, and the Ab3 can neutralize WSSV infection and has Ab1 properties. In the invention, the idiotype antibody is applied in the research of WSSV for the first time; a screening system is established, which uses an indirect enzyme-linked immunosorbent assay (ELISA) method and a competitive enzyme-linked immunosorbent assay (ELISA) method for detection; the fact that Ab3 has properties of Ab1 is proved by adopting an indirect immnnofluotesent method (IIF), a gold labeling immunoelectron microscopic method and crayfish in vivo neutralization tests, thus proving that the monoclonal antibody in the invention has the property to simulate original antigen WSSV-VP28.

Embodiments of the present invention generally relate to proteins derived from white spot syndrome virus, nucleic acid sequences encoding them, and their use in the manufacture of a vaccine for prophylaxis and / or treatment of white spot syndrome in crustaceans.

The invention relates to a shrimp white spot syndrome virus disease detection reagent kit and a detection method thereof. The reagent kit consists of a set of primers, a DNA extracting buffer solution, DNA polymerase, a loop-mediated isothermal amplification reaction liquid, positive contrast, negative contract, betaine and dNTP. The reagent kit can detect whether a shrimp to be detected is infected by a shrimp white spot syndrome virus. The reagent kit utilizes the strand displacement characteristic of Bst DNA enzymes, designs four primers to identify six regions of a target sequence, generates a large number of stem-loop sample structures under the isothermal condition, and does not require any special equipment under the condition of qualitative detection, thereby having the characteristics of specificity, high sensitivity and short detection time compared with common PCR.

The invention relates to a detection kit for isothermal amplification of nucleic acid for the white spot syndrome virus and a detecting method thereof. The kit is designed by using a set of LAMP primers as the main body designed according to the gene conserved sequence of the white spot syndrome virus. The kit is provided with eleven agents needed by WSSV detection including SEMP lapping liquid, nucleic acid extract, UNG enzyme, TE buffer solution, enzymolysis buffer solution, LAMP reaction solution and on the like, thus realizing a programmed and standardized detection; therefore, the invention has the advantages of the highest sensitivity, simplicity, rapidity, safety, good specificity and low cost, and the amount of the copies detected with virus can be as low as 26. Only one water-bath is needed to correctly detect the white spot syndrome virus in prawns and aquaculture water within 2 hours by the kit and the detecting method. Moreover, the problem that the detection is subject to interference in the LAMP technology is also solved. The detecting method of the invention is expected to substitute the previous related detecting methods of the white spot syndrome virus, such as the electron microscope method, the TE staining method, the biopsy method, the antibody detecting method, the nucleic acid probe hybridization method and the PCR detecting method.

The invention discloses an indoor cultivation method of litopenaeus vannamei parent, and relates to the breeding of seawater shrimps. The method disclosed by the invention selects the healthy shrimp seed without carrying white spot syndrome virus, and is characterized in that the parent cultivation is performed in a cement nursery pond in a seed cultivation room; the water change amount of the cement nursery pond is adjusted according to the body length of the shrimp seeds; the shrimp seeds are fed with compound feed before the body length reaches 10cm; the shrimp seeds are fed with the mixture of compound feed and fresh shredded squid when the body length reaches 10cm; when the body length reaches 12cm, the shrimp seeds are fed with fresh shredded squid; after the shrimp seeds are longer than 14cm, the spermatophore of the male shrimp occurs, the shrimp seeds are fed with fresh fait with gonad of clamworm and oyster meat; before the shrimp seeds reach the parent specification, the cultivation salinity is kept at 22; when the shrimp seeds reach the parent specification and the spermatophore of a few male shrimps is mature, the salinity is adjusted to 30; and in the next day after a great amount of water in the nursery pond is changed, 10ml / m<3> of composite bacterial liquid consisting of bacillus subtilis, lactic acid bacteria and saccharomycetes is added once per day.

The present invention relates to a novel composition useful for inhibiting White Spot Syndrome Virus (WSSV) infection of crustacean animals, particularly those of the genera Penaeus sp. More specifically, the novel composition comprises a polypeptide whose amino acid sequence corresponds to at least a portion of Vp28, a surface protein of WSSV, or an antibody that specifically binds the polypeptide. The polynucleotide sequences encoding the Vp28 polypeptides of the present invention are also disclosed. Further disclosed are methods for using the novel compositions to inhibit WSSV infection in crustacean animals.

The invention relates to three-kind of membrane protein of the white spot syndrome virus: VP68,VP281,VP466 having the infective function. Their differential polyclonal antibodies have the ability of resisting the white spot syndrome virus. The invention also deals with the apply of the differential polyclonal antibody of three kinds of membrane protein of the white spot syndrome virus: VP68,VP281,VP466and the producing method of the said membrane protein VP68,VP281,VP466 and their differential polyclonal antibody.

The invention discloses an immunity detection chip of prawn white spot syndrome virus (WSSV), comprising a chip carrier and an agarose gel layer which is paved on the chip carrier, wherein, the agarose gel layer is fixed with a plurality of antibody microarrays of 4*4, and chip-dedicated fences or Super PAP Pen scribings are used for separating the microarrays from each other. The invention comprises the following steps: adopting the sandwich method to detect the antigen; fixing the pathogenic polyclonal antibody (PcAb) on the chip slice base; taking the target organ tissue of the individual to be detected to prepare to-be-detected sample liquid; incubating directly the to-be-detected sample liquid with the chip which is fixed with PcAb to capture the antigen and lead the antigen to combine on the chip; adding a specific monoclonal antibody probe marked by fluorescence; and reading results through a CCD chip scanner. The invention has the advantage of detecting white spot syndrome virus (WSSV) in multiple samples simultaneously, and is applicable to the rapid and accurate detection of white spot syndrome virus (WSSV) of prawns / crabs in breeding production and the quarantine inspection of WSSV in import and export prawns / crabs.

The technology belongs to the technical field of aquiculture disease prevention, in particular to the preparation method of the immune reinforcing agent for cultivating crustacean, and the applicationmethod thereof. The aquiculture composite immune reinforcing agent comprises 2-4 parts of aquiculture extractive or baicalin and 2.5-5 parts of levamisole. The composite immune reinforcing agent caneffectively enhance non-specific immunity for cultivating blue crab, improve the activity of the blood serum phenol oxidase, the superoxide dismutase and the alkalinephosphatase of the blue crab, andimproves the disease resistance of the blue crab to common pathogens such as white spot syndrome virus, vibrio, hematodinium, etc. The composite immune reinforcing agent can be applied to the cultivation of the blue crab by being injected or added into blue crab fresh fish bait and granulated feed for preventing and controlling the disease at the disease peak and reducing the disease rate and themortality rate of the blue crab. The immune reinforcing agent is added into the feed for preventing and controlling the disease of the penaeus vannamei boone and exopalaemon carinicauda.

The invention discloses a main cis-acting element of a shrimp white spot syndrome virus (WSSV) iel promoter and a transcription factor combined with the same and application. In the invention, by starting from transcriptional regulation of WSSV iel and carrying out structural and functional analysis on the promoter of the WSSV iel through deletion and mutation, a 12-bp DNA is found to be the maincis-acting element of the WSSV iel and is a crucial factor for the high expression of the iel. In the invention, a DNA affinity chromatography method is used for purifying a protein combined with a DNA segment from a nucleus protein Sf9, the protein is identified to be PHB2 (Poly-Beta-Hydroxybutyrate 2) through biological mass spectrometry, and the interaction between the protein and the DNA segment is proved to be specific by an electrophoretic mobility shift assay. Experiment results prove that PHB2 serves as a transcription factor and is specifically combined with a 12-bp DNA sequence of the iel promoter to start WSSV immediate early gene transcription so as to further regulate the replication of the WSSV, and can be used as an effective action target of medicaments for screening medicaments for resisting the shrimp WSSV.