67 results about "Streptococcus iniae" patented technology

The invention provides bacillus velezensis and application of the bacillus velezensis as an aquatic pathogenic bacteria inhibitor. The bacillus velezensis is separated from the intestinal canal of healthy tilapia, and the preservation number of the bacterial strain is GDMCC No. 60344. The bacillus velezensis has a broad-spectrum antagonistic aquatic common pathogenic bacteria function, and has theadvantages that the growth of the pathogenic bacteria, such as streptococcus agalactiae, streptococcus iniae,nocardia seriolea, aeromonas hydrophila, aeromonas schubertii and edwardsiella tarda, canbe effectively inhibited, the use is safe, no residue is generated, reducing the usage amount and the residual amount of chemical drugs in aquaculture is facilitated, the production cost is reduced, and the practicality is strong.

The invention discloses high yield and high efficiency health culture feed of tilapia and a preparation method thereof. The feed comprises, by weight, 30 to 50 portions of protein powder, 10 to 18 portions of soybean meal, 6 to 12 portions of peanut meal, 8 to 14 portions corn, 4 to 7 portions of cottonseed meal, 5 to 8 portions of rapeseed meal, 10 to 16 portions of high-gluten flour, 1.8 to 3 portions of vitamins, 1 to 2 portions of amino acids, 1 to 1.8 portions of trace elements, 5 to 8 portions of vegetable oil, 1 to 2 portions of an attractant and 0.6 to 0.9 portions of a dolphin streptococcal biological preparation. The feed is prepared through drying, pulverizing, mixing and drying. The feed has a reasonable formula, is rich in nutrients, can increase the feed intake of tilapia suffering from streptococcal diseases, increases the growth rate, enhances the immune functions, reduces the death rate of tilapia suffering from streptococcal diseases and improves the cure rate of the streptococcal diseases.

The invention relates to the field of the molecular immunology, in particular to a streptococcus iniae vaccine and preparation and application thereof. Particularly, the vaccine is expressed by a base sequence in a sequence table SEQ ID No.1. A preparation method comprises the following steps: using streptococcus iniae G26 as a template and using a primer F2 / R5 to carry out PCR; and connecting a PCR product with plasmids pBTA1 and converting colon bacillus DH5l alpha by the connecting solution to obtain a bacterial strain DH5 alpha / pTASP11, i.e. a bacterial strain of the streptococcus iniae vaccine, which expresses a protein encoded by the base sequence in the sequence table SEQ ID No.1. The application of the streptococcus iniae vaccine is that the obtained bacterial strain of the streptococcus iniae vaccine is cultured by an LB culture medium until OD600 is 0.8 and then is dissolved in PBS and the obtained vaccine preparation solution has effect on immune protection for the streptococcus iniae. The obtained vaccine of the invention has very simple preparation process and does not need any extraction and purification process. The protection efficiency of the streptococcus iniae vaccine aiming at streptococcus iniae infection reaches 100 percent.

The invention provides a triple real-time fluorescent PCR method and a kit for detecting three streptococci at the same time. The kit comprises: a streptococcus agalactiae detection primer pair which is represented as the Seq ID No.1 and the Seq IN No.2 in the sequence table, a probe which is represented as the Seq ID No.3 in the sequence table, a streptococcus dysgalactiae detection primer pair which is represented as the Seq ID No.4 and the Seq ID No.5 in the sequence table, a probe which is represented as the Seq ID No.6 in the sequence table, a streptococcus iniae detection primer pair which is represented as the Seq ID No7 and the Seq ID No.8 in the sequence table, a probe which is represented as the Seq ID No.9 in the sequence table, three positive comparison sample which are represented as the Seq ID No.16, the Seq ID No.17 and the Seq ID No.18 in the sequence table, a negative comparison sample (ddH2O), a PCR buffering liquid required in fluorescent real-time quantification PCR amplification, dNTP and TaqDNAPolymerase. The method is simple and quick and is good in specificity.

The invention discloses a method for improving the streptococcus iniae infection-resisting capacity of GIFT oreochromis niloticus during high-temperature stress. The method comprises the following steps: preparing basic ration and a compound Chinese medicinal herb formula, and feeding feed. Due to the adoption of the method, the immunity index of the GIFT oreochromis niloticus during high-temperature stress can be effectively increased, the liver damage is reduced, meanwhile, the streptococcus iniae infection-resisting capacity of the GIFT oreochromis niloticus can be improved, the morbidity is reduced, and the survival rate of the GIFT oreochromis niloticus infected with streptococcus iniae can be increased. Several Chinese medicinal herbs involved in the method are relatively available and relatively low in cost; a production technology is simple. The basic ration with the compound Chinese medicinal herbs has relatively good palatability, and after the basic ration is taken by the GIFT oreochromis niloticus, drug residues in bodies of the GIFT oreochromis niloticus are relatively few, and therefore, the GIFT oreochromis niloticus meets the requirements on green healthy aquatic products.

The invention discloses a phage flocculant and application of the phage flocculant in a fermentation post-treatment technology and belongs to the field of a microbial post-treatment technology. The phage flocculant is characterized in that the phage flocculant is a cationic flocculant, and a working concentration of the phage flocculant is 0.1-5% generally. The flocculant is applied in the fermentation post-treatment technology of a vibrio parahaemolyticus phage, a coliphage, a salmonella phage, a xanthomonas axonopodis phage, a ralstonia solanacearum phage, a staphylococcus aureus phage, a streptococcus agalactiae phage, a streptococcus iniae phage or a pseudomonas aeruginosa phage. The phage flocculant can play a role of rapid subsidence of bacteria, be convenient and quick to operate, reduce centrifugal equipment investment and manpower and material resource loss and meet requirements of industrial mass production of the phage, and is safe, nontoxic and harmless to animals and plants.

The invention discloses a diagnostic kit for a Streptococcus iniae molecule and a detection method. The kit comprises lysis buffer solution A, DNA extraction B solution, DNA extraction C solution, DNA extraction D solution, positive template contrast E solution and PCR reaction solution F solution. The kit and the detection method consist of a pair of specific primers designed by a conserved sequence in a Streptococcus iniae gene group as a main body. Meanwhile, the invention optimizes the DNA extraction lysis solution and PCR reaction conditions, including magnesium ion concentration, annealing temperature and recurrent number. The kit can carry out qualitative detection on a specific DNA fragment of the Streptococcus iniae, is simple, convenient and rapid, has good specificity and high sensitivity, can be used for bacterium tracking detection of each cultivating process for fish cultivation, and has higher practical value.

The invention relates to the technical field of gene engineering, in particular to novel broad-spectrum chimeric lysin BGS-PlySb and an encoding gene and application thereof. The novel broad-spectrum chimeric lysin BGS-PlySb has an amino acid sequence shown as in SEQ ID NO. 1; the encoding gene of the novel broad-spectrum chimeric lysin BGS-PlySb has a nucleotide sequence shown as in SEQ ID NO. 2. A novel chimeric lysin is constructed by means of gene splicing and is suitable for killing various species of Streptococcus and Staphylococcus, particularly various species of Enterococcus, and Staphylococcus aureus, as well as Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus suis, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus mutans, Streptococcus iniae and other species; the novel broad-spectrum chimeric lysin BGS-PlySb has good stability and is insensitive to high-concentration NaCl, recombinant protein GBS-PlySb is well expressible in Escherichia coli BL21 (DE3), and a high dose of GBS-PlySb is free of significant cytotoxicity. Therefore, the novel broad-spectrum chimeric lysin BGS-PlySb is applicable independently to or as an additive to the control of various species of Streptococcus and the treatment of infections caused by these species, and has a promising application prospect.

The present invention discloses a feed for preventing and curing streptococcus iniae diseases of oreochromis mossambicus and a preparation method thereof. The feed comprises the following raw materials in parts by weight: 30-50 parts of protein powder, 3-5 parts of beet pulp, 4-6 parts of earthworm powder, 12-18 parts of sweet potatoes, 4-7 parts of cottonseed meal, 5-8 parts of rapeseed meal, 9-14 parts of flour, 1.8-3 parts of vitamins, 1-2 parts of amino acids, 1-1.8 parts of trace elements, 5-8 parts of vegetable oil, 1-2 parts of an attractant and 0.6-0.9 part of a streptococcus iniae biological preparation. The feed is prepared by the steps of sun-drying, crushing, mixing, drying, etc. The feed is reasonable in preparation and rich in nutrition, can increase the food intake amount of the oreochromis mossambicus suffering from the streptococcus iniae diseases, increases the growth speed, enhances the body immune functions, reduces the death rate of the oreochromis mossambicus suffering from the streptococcus iniae diseases and improves the cure rates.

The invention discloses a method for improving the streptococcus iniae infection resisting capability of GIFT tilapia in high-density culture. The method comprises the steps of preparing of compound Chinese herbal medicine, adding, feeding and breeding management. Compared with the prior art, the method has the following advantages: (1) the immunological protection of the GIFT tilapia in high-density culture can be effectively improved, and the survival rate of GIFT tilapia infected with streptococcus iniae is improved; (2) the compound Chinese herbal medicine added in a feed belongs to environment-friendly feed additives, and has less negative effect on fish and water; (3) the blood and head-kidney immunological stress response can be effectively enhanced, and the liver stress injury is reduced; (4) the adopted technological steps of the method are simple, the Chinese herbal medicine is low, and easy to obtain and preserve, and can be well absorbed and utilized by a fish body, and excrement of the GIFT tilapia does not impact the water quality.

The invention discloses a detection primer group, a detection kit, and a detection method of nocardia seriolae, vibrio harveyi, and streptococcus iniae. The detection primer group comprises a primer pair Ns-mce, a primer pair Vh-tox, and a primer pair Si-its; the nucleotide sequences of the primer pairs are represented by SEQ ID NO.1-6. The detection kit comprises above detection primer group. According to the detection method, the detection kit is used for detection; samples are subjected to pretreatment, genome DNA is extracted as sample group DNA, and PCR is adopted to detect the samples. A triplex PCR detection method is established for tracking measurement of nocardia seriolae, vibrio harveyi, and streptococcus iniae at different periods, avoiding transmission of pathogenic bacteria, and improving rapid inspection and quarantine of pathogenic bacteria in aquatic products; and the invention belongs to the field of aquatic cultured animal pathogenic bacteria detection.

The present invention discloses primers and a probe for streptococcus iniae LAMP-LFD visual detection, and an application of the primers and the probe. According to the present invention, three pairs of LAMP primers such as gyrB-F3, gyrB-B3, gyrB-FIP, gyrB-BIP, gyrB-LF and gyrB-LB, and a probe gyrB-HP are provided, and the nucleotide sequences are represented by SEQID NO.1-EQID NO.7; and with the primers and the probe, a LAMP reaction system amplification step, a LAMP reaction product probe hybridization step and a LFD detection step are performed so as to achieve streptococcus iniae visual detection and further be used for kit preparation, wherein advantages of high rapidness, high specificity and high sensitivity are provided, instrument requirements are simple, early diagnosis and detection of streptococcus iniae can be easily achieved, and requirements of grass-roots detection facilities and field epidemic focus detection.

Disclosed is a vaccine for edwardsiellosis and streptococcosis in a fish. Specifically, disclosed is a vaccine for edwardsiellosis in a fish, which comprises inactivated cells of (A) an Edwardsiella tarda strain derived from the target fish, and inactivated cells of (B) an Edwardsiella tarda strain derived from a fish other than the target fish, wherein, when the strain (A) is a typical Edwardsiella tarda strain, the strain (B) is an atypical Edwardsiella tarda strain, whereas when the strain (A) is an atypical Edwardsiella tarda strain, the strain (B) is a typical Edwardsiella tarda strain. Also specifically disclosed is a vaccine for edwardsiellosis and / or streptococcosis in a fish, which comprises the components mentioned above and inactivated cells of (C) Streptococcus iniae and / or Streptococcus parauberis.

The invention discloses Bacillus amyloliquefaciens SCAU-070 and application thereof. The Bacillus amyloliquefaciens SCAU-070 is obtained by separating and screening, the strain is deposited in the Guangdong Microbial Culture Collection Center on July 1, 2020, and the collection number is GDMCC No: 61074. The strain can significantly promote the growth of aquatic animals, inhibit the growth of aquatic animal pathogenic bacteria such as streptococcus agalactiae, streptococcus iniae, micrococcus luteus and vibrio parahaemolyticus in aquatic animal breeding water environment, improve the antioxidant ability of aquatic animal tissues, and regulate the content of cytokines in aquatic animal tissues; and therefore, the strain is of great significance in reducing the use of antibiotics and prohibited drugs, and has good application prospects and market value in the preparation of aquatic animal feed additives, microecological preparations for regulating the aquatic animal breeding water environment and medicines for improving the body immunity of aquatic animals.

Isolated M proteins, or M-like proteins, of Streptococcus iniae, encoding nucleic acids, genetic constructs and antibodies which bind the isolated M proteins are provided. Also provided are isoforms and / or fragments of the M proteins, or M-like proteins, that display reduced fibrinogen binding. The isolated M proteins, antibodies and encoding nucleic acids maybe useful for diagnosis, immunization and / or therapy of Streptococcus iniae infections of animals such as fish and humans.

The invention relates to the field of vaccinology, in particular to a Streptococcus iniae trivalent DNA vaccine and to a construction method thereof. The method comprises the steps of using Streptococcus iniae G26 as a template, allowing the primers of F1 / R1, F2 / R2 and F3 / R3 to carry out PCR amplification respectively, allowing the product and a vector pCN3 plasmid to be subjected to connection and transformation, and allowing the plasmid to be subjected to equivalent volume mixing to obtain the trivalent DNA vaccine. The efficiency of the protection of the vaccine obtained by the invention against Streptococcus iniae infection is more than 90% in one month or two months after immunization.

The present disclosure relates to genetic markers for discrimination and detection of bacteria causing Edwardsiellosis and Streptococcosis in fish, and a method for discriminating and detecting the bacteria using the same. A genetic marker for discrimination and / or detection of each of Edwardsiella tarda, Streptococcus iniae, Streptococcus parauberis and Lactococcus garvieae, which cause fish diseases, is selected and a peptide nucleic acid and a primer pair, which are specific for the genetic marker, are used to amplify and obtain melting curves having different fluorescences depending on bacterial species. Thus, bacteria that cause fish diseases can be discriminated and whether or not fish would be infected with the bacteria can be detected in a simple, rapid and accurate manner.

The invention discloses a streptococcus iniae loop-mediated isothermal amplification kit and application thereof. The kit comprises LAMP primers, a 2*reaction buffer, a Bst DNA polymerase, a fluorescent visual detection reagent, ultra-pure water and a streptococcus iniae DNA template, wherein the LAMP primers comprise outer primers F3 and B33 and inner primers FIP and BIP. Specificity detection and sensitivity detection show that the LAMP detection method provided by the invention can be used for monitoring reaction in real time and detecting the copy number of streptococcus iniae quantitatively so that the detection result can be obtained rapidly and accurately and convenience is brought for simple and rapid detection of the streptococcus iniae.

The present invention relates to a field of vaccinology, and especially relates to a bacteria-delivered cross protection vaccine against Edwardsiella tarda and Streptococcus iniae. The cross protection vaccine is a recombinant strain obtained by plasmid [rho]DKS10 transforming escherichia coli DH5[alpha]. The vaccine obtained in the present invention has 83% and 75% protection efficiency respectively for Edwardsiella tarda and Streptococcus iniae infection.

The invention discloses a molecular marker and a screening method of gift strain nile tilapia oreochromis niloticus streptococcus iniae infection resistant family. The molecular marker is a sequence shown in SEQ ID NO.20; the screening method comprises five steps of streptococcus iniae infection, genome DNA extraction, primer screening, SRAP-PCR (Sequence-Related Amplified Polymorphism-Polymerase Chain Reaction) amplification and sequencing and analyzing, by the method, a parent family with stronger resistance to streptococcus iniae disease can be found more accurately and quickly, after enhanced cultivation, the parent family can be used as the parent for fingerlings propagation. The molecular marker screened out is prepared into a probe for detecting different parent families, the probe can sort out the streptococcus iniae disease resistant parent more simply and accurately, and selection of parent among different families is facilitated. Meanwhile, by using the bred disease-resistant parent as the propagation parent, quality of fingerlings can be improved further, and culture benefit of gift strain nile tilapia oreochromis niloticus can be increased.

The invention relates to the field of molecular biology, in particular to an application of serine protease (PoCFI-Tryp) of a flounder complement regulatory protein I factor. The invention relates tothe application of serine protease (PoCFI-Tryp) in the flounder complement regulatory protein I factor in preparation of a bacteriostatic agent. The serine protease disclosed by the invention can be combined with various bacteria, and can obviously inhibit the growth of Edwardsiella tarda, Vibrio anguillarum, Vibrio harveyi and Streptococcus iniae. The obtained protein has the application potential in bacterial infection resistance.

The invention discloses polypeptide shown in the amino acid sequence SEQ ID NO:1 and further discloses a purpose of polypeptide in preparing an immunologic adjuvant of a vaccine. The invention further discloses the immunologic adjuvant prepared from polypeptide and an adjuvant. By means of IL-8 polypeptide, the immunoprotection effect of the vaccine especially a streptococcus iniae subunit vaccine can be remarkable enhanced, and polypeptide can be used as the adjuvant of the vaccine. Polypeptide is safe to use, easy to prepare and convenient to use, and new effective choices are provided for prevention and treatment of fish diseases.

The invention provides a paralichthys olivaceus streptococcus iniae GAPDH series-connection multi-epitope polypeptide and application thereof. The provided series-connection multi-epitope polypeptideis formed by binding four antigen epitope peptides through peptide joints, wherein the amino acid sequences of the four antigen epitope peptides are shown in SEQID NO:1-4 respectively. One specific amino acid sequence of the series-connection multi-epitope polypeptide is SEQID NO:8. The invention further provides a streptococcus iniae GAPDH multi-epitope vaccine prepared by using the series-connection multi-epitope polypeptide as an antigen and a vaccine adjuvant. The multi-epitope vaccine can provide strong immune protection for paralichthys olivaceus resistance to S.iniae infection, and theeffect is significantly higher than that of a GAPDH recombinant protein subunit vaccine and a S.iniae inactivated vaccine.

The invention relates to a primer combination capable of detecting three kinds of streptococcus simultaneously. The primer combination consists of upstream and downstream primers capable of detectingstreptococcus agalactiae, as shown in SEQ ID MO:1, 2, upstream and downstream primers capable of detecting streptococcus dysgalactiae, as shown in SEQ ID MO:3, 4, and upstsream and downstream primerscapable of detecting streptococcus iniae, as shown in SEQ ID MO:5, 6. A reagent kit consists of a DNA extraction reagent and a PCR reaction reagent, wherein the DNA extraction reagent consists of a TEbuffer solution, proteinase k, chloroform, isopropyl alcohol, ethanol and redistilled water, and the PCR reaction reagent contains a reaction buffer solution, dNTPs, DNA polymerase, the 6 primers anddeionized water. A method for detecting the three kinds of streptococcus simultaneously comprises the steps of extracting DNA in samples through the extraction reagent, performing PCR amplification on the DNA through the reaction reagent, and performing gel electrophoresis on amplification products. The three kinds of streptococcus can be detected simultaneously, detection is simple, quick, highin sensitivity and high in specificity, and the primer combination and the method can meet requirements for large-scale molecular epidemiology investigation and analysis of aquatic animals and qualitysafety detection of aquatic products.

Isolated M proteins, or M-like proteins, of Streptococcus iniae, encoding nucleic acids, genetic constructs and antibodies which bind the isolated M proteins are provided. Also provided are isoforms and / or fragments of the M proteins, or M-like proteins, that display reduced fibrinogen binding. The isolated M proteins, antibodies and encoding nucleic acids maybe useful for diagnosis, immunization and / or therapy of Streptococcus iniae infections of animals such as fish and humans.

The invention discloses a primer for performing double-PCR early and rapid detection on streptococcus agalactiae and streptococcus iniae as well as application thereof. A sample is detected by utilizing the primer disclosed by the invention, and streptococcus agalactiae and streptococcus iniae in the ample can be simultaneously specially detected at one time. A detection method built by the invention can be used for detecting the streptococcus agalactiae of which gene group content is 9.84x10<-5>ng / microliter and the streptococcus iniae of which gene group content is 9.30x10<-5>ng / microliter,and the streptococcus agalactiae with bacteria liquid concentration of 2.76x102cfu / mL and the streptococcus iniae bacteria liquid concentration of 2.51x102cfu / mL. The method disclosed by the invention has the advantages that the method is higher in specificity, sensitivity, repeatability and stability on quickness and correctcness in a detection process of tilapia samples, early molecular warningof tilapia streptococcus diseases can be realized, the application prospect is wide, and an accurate and effective mean is provided for quick diagnosis of the tilapia streptococcus diseases and investigation of epidemiology.

The invention relates to the molecular biology and immunology field, in particular to a gram positive microbes DNA vaccine and construction and application thereof. The vaccine has base sequence in sequence table SEQ ID No.1. The preparation method is that an eukaryotic expression vector pCN3 is used to construct streptococcus iniae DNA vaccine plasmid pCNS 10 which is suspended in PBS to be the DNA vaccine. The DNA vaccine has vaccine protection effect for streptococcus iniae and is the first construction in the whole world. The application is simple and once vaccine can achieve high efficiency and long-term vaccine protection effect.

The invention discloses a streptococcus iniae disease biological agent and a preparing method and application thereof, and belongs to the technical field of prevention and treatment of the tilapia streptococcus iniae disease. The streptococcus iniae disease biological agent is prepared through fermentation tank empty sterilization, culture medium preparation, culture medium sterilization, strain inoculation, strain fermentation, concentration, drying, smashing and screening. By adding the streptococcus iniae disease biological agent to a feed, the immunity and anti-stress capacity of tilapia can be effectively improved, the death rate of tilapia from the streptococcus iniae disease can be reduced, the cure rate can be increased, and productivity effectiveness can be improved.

The GapC plasmin binding protein genes of Streptococcus dysgalactiae (S. dysgalactiae), Streptococcus agalactiae (S. agalactiae), Streptococcus uberis (S. uberis), Streptococcus parauberis (S. parauberis), and Streptococcus iniae (S. iniae) are described, as well as the recombinant production of the GapC proteins therefrom. Also described is the use of the GapC proteins from those species in vaccine compositions to prevent or treat bacterial infections in general, and mastitis in particular.

The GapC plasmin binding protein genes of Streptococcus dysgalactiae (S. dysgalactiae), Streptococcus agalactiae (S. agalactiae), Streptococcus uberis (S. uberis), Streptococcus parauberis (S. parauberis), and Streptococcus iniae (S. iniae) are described, as well as the recombinant production of the GapC proteins therefrom. Also described is the use of the GapC proteins from those species in vaccine compositions to prevent or treat bacterial infections in general, and mastitis in particular.