35 results about "Streptococcus mastitidis" patented technology

The invention relates to the technical field of biology, in particular to an SNP (Single Nucleotide Polymorphism) marker combination for identifying tilapia streptococcus agalactiae disease resistance and application thereof. The invention discloses 11 SNP loci significantly related to streptococcus agalactiae resistance in a tilapia UTRN gene, and constructs 18 SNP marker combinations for streptococcus agalactiae resistance phenotype identification, so that the resistance phenotype identification accuracy and hereditary stability can be effectively improved. Besides, according to the SNP typing primer and method based on the # imgabs0 # MALDI-TOF mass spectrum platform, high-throughput and high-precision genetic typing can be achieved, and the SNP typing primer and method have important application value in resistance phenotype identification and assistant selective breeding.

The application discloses a kind of fluorescence colorimetric double-mode detection streptococcus agalactiae without aerosol pollution method, comprising the following steps: (1) strain culture and DNA extraction;(2) the combination of UDG and LAMP;(3) UDG-LAMP-CRISPR fluorescence mode detection system;(4) colorimetric mode detection system: read image RGB value in G value, G value significantly increases, and it is positive result.This patent innovatively combines UDG, LAMP, CRISPR and G4, introduces connection probe to realize "fluorescence-colorimetric" single system double-mode detection.The two detection modes of the detection platform can be freely split or combined, and can meet the needs of different detection environments.By combining UDG system and LAMP technology, aerosol pollution possibly carried in amplification process is avoided.Connection probe is introduced, CRISPR fluorescence detection is combined with G4 colorimetric detection, a detection system with "fluorescence-colorimetric" two detection modes is established, which is used for quantitative identification and cross verification of streptococcus agalactiae.

The application discloses a bocachito Nat10 antibacterial peptide, a coding sequence and use and a preparation method thereof, and relates to a method based on multiomics, a biological big data based on multiomics data of bocachito, screening and identifying an antibacterial peptide precursor gene sequence, and obtaining an amino acid sequence of the bocachito Nat10 antibacterial peptide through translation design, as shown in SEQ ID NO: 3. Through activity simulation prediction and in-vitro experiment verification, the bocachito Nat10 antibacterial peptide shows inhibitory effects on the growth activities of Streptococcus agalactiae, Salmonella enteritidis, Staphylococcus aureus, Vibrio anguillarum and Escherichia coli, especially on Staphylococcus aureus, and shows the best antibacterial effect at a concentration of 2.5 micromoles per liter. The bocachito Nat10 antibacterial peptide can be used for preparing various bactericidal preparations, including bacteriostatic agents, medicines or additives, and has good practical value and social ecological benefits.

The present application relates to the field of biotechnology, and particularly relates to a KASP molecular marker related to the resistance of tilapia to streptococcus agalactiae disease and an application thereof.Two SNP sites (SNP-1 and SNP-2) significantly associated with the resistance are screened from the intron region of EGFRX2 gene of tilapia, a typing method of the above sites is developed through KASP technology, and the correlation between the SNP-1 genotype and the resistance phenotype is verified.Further, an advantage haplotype individual with the resistance to streptococcus agalactiae disease is screened by combining the two SNP sites, and the survival rate of the individual in the challenge experiment is significantly improved.The detection kit provided by the present application can realize high-throughput and high-precision typing, is suitable for the identification of the resistance genotype of tilapia and the marker-assisted selection, provides an efficient genetic improvement tool for the disease-resistant breeding of tilapia, and is helpful to reduce the loss of breeding diseases and reduce the dependence on antibiotics.

The invention discloses a streptococcus agalactiae attenuated strain, a vaccine and application of the streptococcus agalactiae attenuated strain, and belongs to the technical field of veterinary biological products. The streptococcus agalactiae attenuated strain is obtained by knocking out a marR gene of a streptococcus agalactiae virulent strain HN016; wherein the nucleotide sequence of the marR gene is as shown in SEQ ID NO: 1. After the marR gene of the streptococcus agalactiae virulent strain HN016 is knocked out, the streptococcus agalactiae attenuated strain of which the virulence is remarkably reduced is obtained, and after the streptococcus agalactiae attenuated strain is prepared into the vaccine, the survival rate of aquatic animals infected with streptococcus agalactiae can be remarkably improved, so that the survival rate of the aquatic animals infected with streptococcus agalactiae is improved. The streptococcus agalactiae attenuated strain has a good application prospect in preparation of drugs for preventing and / or treating streptococcus agalactiae infection of aquatic animals.

The application discloses a recombinant strain and a preparation method thereof and application of the recombinant strain in preparation of a vaccine for Streptococcus agalactiae of tilapia, and the recombinant strain is successfully constructed by the following steps: designing an ORF4 primer, amplifying an ORF4 gene fragment from a DNA template of Streptococcus agalactiae, editing the ORF4 gene fragment into Escherichia coli by means of a CRISPR / Cas9 gene editing technology, and connecting a green fluorescent protein (GFP). The recombinant strain is subjected to immunoprotection research on the Streptococcus agalactiae of tilapia, and it is shown that the recombinant strain has a good protection effect, and can provide a solid theoretical basis and technical support for application and development of the vaccine for Streptococcus agalactiae of tilapia in the field of aquaculture.

The present application relates to the field of biotechnology, and more particularly to a SNP marker related to the level of anti-streptococcus agalactiae IgM antibody of tilapia and application thereof. The present application screens out QTLs, functional genes and key signal pathways significantly related to the level of anti-streptococcus agalactiae IgM antibody of tilapia by RNA-Seq and BSA-Seq technologies, and further identifies two SNP markers closely related to the level of anti-streptococcus agalactiae IgM antibody of tilapia, thereby providing important genetic targets and tools for molecular marker-assisted breeding of anti-streptococcus agalactiae of tilapia.

The invention belongs to the field of microorganisms, and discloses a bacterial strain having an antagonistic effect on pathogenic bacteria of cow mastitis and application of the bacterial strain, the bacterial strain SUD-8 having the antagonistic effect is paenibacillus polymyxa, and the bacterial strain SUD-8 has the antagonistic effect on the pathogenic bacteria of cow mastitis and application of the bacterial strain SUD-8 to the pathogenic bacteria of cow mastitis. The strain has an antagonistic effect on klebsiella pneumoniae, escherichia coli, staphylococcus aureus, streptococcus uberis and streptococcus dysgalactiae, and the growth speed in the cow dung back-mattress material is high.

The present application relates to the field of biotechnology, and more particularly to a detection method for resistance of tilapia to streptococcosis, a KASP primer set and application. The present application identifies a key SNP site of the ABCB4 gene significantly associated with resistance of tilapia to streptococcosis by the method of population genetics, and develops a genotyping method for the SNP site by KASP technology, thereby realizing efficient evaluation of the disease-resistant trait. Experimental data show that the survival time of AA genotype individuals in the challenge test is longer than that of GA / GG genotype individuals, the proportion of resistant fish is significantly improved, and the genotype frequency distribution is strongly correlated with the disease resistance of the population. The breeding strategy based on the SNP marker can improve the overall disease resistance of the breeding population and reduce the mortality rate, thereby providing an efficient genetic improvement tool for breeding of tilapia resistant to streptococcosis, and helping to reduce the loss of breeding diseases and reduce the dependence on antibiotics.

The present application relates to the field of microbial technology, in particular to bacillus subtilis BS-1 and its application in water buffalo silage feed, the strain BS-1 of the present application has good bacteriostatic activity on escherichia coli, salmonella and streptococcus uberis, after mixing and fermenting according to a specific proportion with orange peel and cassava residue, the bacteriostatic spectrum of the obtained silage feed is significantly enhanced, which can simultaneously inhibit escherichia coli, campylobacter jejuni, salmonella, streptococcus uberis, staphylococcus aureus and streptococcus agalactiae that cause diarrhea and mastitis. In the feed, after mixing 60%-75% of orange peel and 25%-40% of cassava residue, the crude protein content of the fermentation product is significantly improved, and the fiber content is reduced. Feeding test shows that the feed can significantly improve the milk yield and milk quality of lactating water buffalo, and effectively alleviate diarrhea. The present application provides a safe, efficient and nutritious silage feed with disease prevention function.

The invention discloses a lactobacillus plantarum bacteriostatic agent and application thereof, and relates to the technical field of microbial engineering. According to the lactobacillus plantarum LP2301 provided by the invention, fermentation liquor of the lactobacillus plantarum LP2301 can be used for effectively inhibiting aquatic pathogenic bacteria streptococcus agalactiae, limiting the growth of the streptococcus agalactiae, reducing the use of antibiotics, ensuring food safety and guaranteeing the health of animals and human beings by secreting metabolites such as bacteriostatic proteins and organic acids. Secondly, the invention provides an application of the lactobacillus plantarum bacteriostatic agent, and the lactobacillus plantarum LP2301 is compounded into the bacteriostatic agent; through multi-target cooperation of the cortex phellodendri extract and the rosmarinic acid, the antibacterial efficiency of single probiotics is remarkably optimized; active ingredients of cortex phellodendri directionally destroy a biological membrane barrier of streptococcus agalactiae and disintegrate an adhesion defense system of pathogenic bacteria; rosmarinic acid blocks a bacterial energy metabolism pathway synchronously, so that pathogens lose proliferative power; and exclusive bacteriocin secreted by the LP2301 accurately breaks down a cell membrane structure, so that fatal killing is realized.

The application belongs to the technical field of Chinese herbal medicines, and particularly relates to a Chinese herbal medicine composition for resisting Streptococcus agalactiae, a preparation method and application thereof. Raw materials for preparing the Chinese herbal medicine composition include cortex mali and cumin. The Chinese herbal medicine composition is easy to obtain and simple to prepare, has no chemical residues, is not prone to drug resistance, has significant bacteriostatic activity on Streptococcus agalactiae, and can be applied to the prevention and treatment of Streptococcus agalactiae disease of fish such as tilapia.

The present invention relates to molecular biology and aquaculture breeding, specifically to a method for obtaining a PRMT2 gene mutant by knocking out the gene based on CRISPR / Cas9 technology, and establishing an antibacterial strain of tilapia with a PRMT2 knockout. By knocking out this gene, a tilapia strain resistant to Streptococcus agalactiae can be obtained. Infection experiments revealed that tilapia with a PRMT2 knockout gene exhibited enhanced antibacterial activity compared to wild-type tilapia. This invention provides an important model for disease-resistant tilapia breeding and offers gene targets and new research directions for creating more disease-resistant fish strains and varieties.

The present invention relates to compositions and methods for detecting group B streptococcus (GBS, Streptococcus agalactiae) and identifying the most common genes responsible for clindamycin resistant GBS, such as ermB, ermTR, ermT, lsaC and lsaE, by multiplex real-time PCR assays.

The invention discloses a condom added with sophocarpidine hyaluronic acid lubricating liquid and a preparation method of the condom, and belongs to the technical field of condom production. The core of the method is that after matrine is dissolved, the matrine is added into hyaluronic acid lubricating liquid for the condoms, the hyaluronic acid lubricating liquid containing the matrine is prepared through stirring and mixing, the concentration of the matrine in the hyaluronic acid lubricating liquid is 7.7-25.0 mg / g, when the condoms are subjected to inner packaging, the matrine-containing lubricating agent is added into the condoms, and when the condoms are subjected to inner packaging, the matrine-containing lubricating agent is added into the condoms. And preparing the condom added with the sophocarpidine-containing hyaluronic acid lubricating liquid. According to the condom, inflammation generated after vaginal epithelial cells are stimulated by bacterial structural components is improved through matrine, and the common pathogenic bacteria such as staphylococcus aureus, escherichia coli, enterococcus faecalis and streptococcus agalactiae of vaginitis are inhibited in the using process; and the up-regulation level of NF-kB phosphorylation generated after lipopolysaccharide and peptidoglycan stimulate vaginal epithelial cells is relieved.

The invention relates to immunogenic compositions comprising a capsular polysaccharide (CP) from Streptococcus agalactiae, commonly referred to as group B streptococcus (GBS), conjugated to a carrier protein, and optionally including an aluminum-based adjuvant. The invention further relates to methods for inducing an immune response in subjects against GBS and / or for reducing or preventing invasive GBS disease in subjects using the compositions disclosed herein. The resulting antibodies can be used to treat or prevent GBS infection via passive immunotherapy.

The application discloses a tilapia Npy antibacterial peptide, a coding sequence, and a use and a preparation method thereof, and relates to the following technical scheme: based on a multi-omics method, an antibacterial peptide precursor gene sequence is screened and recognized from a tilapia multi-omics data set based on biological big data, and then the tilapia Npy antibacterial peptide amino acid sequence is obtained through translation design, as shown in SEQ ID NO: 3; through activity simulation prediction and in-vitro experiment verification, the tilapia Npy antibacterial peptide shows inhibitory activity to gram-positive streptococcus agalactiae, staphylococcus aureus and gram-negative escherichia coli, especially to streptococcus agalactiae and staphylococcus aureus, and shows good antibacterial effect at a low concentration (2.5 μmol / L), and can be used for preparing antibacterial products, including preparation of an antibacterial agent, preparation of a medicine or preparation of an additive, and has good practical value and social ecological benefits.

The present application relates to oligosaccharide composition and its use. The present application particularly relates to the use of oligosaccharide composition in the preparation of products for preventing and / or treating aquaculture diseases caused by Vibrio parahaemolyticus, Vibrio harveyi and Streptococcus agalactiae, wherein the oligosaccharide composition is obtained by degrading polygalacturonan or pectin by a polygalacturonase as shown in SEQ ID No: 1, under the conditions of a polygalacturonase concentration of 0.045 mg / mL to 0.055 mg / mL, a polygalacturonan concentration of 7% to 9%, and a degradation time of 12 hours to 20 hours. The present application also relates to the use of oligosaccharide composition in aquaculture. The oligosaccharide composition of the present application has good bacteriostatic effect and disease prevention and control effect in aquaculture.

The present invention relates to oligosaccharide compositions and uses thereof. The invention particularly relates to application of an oligosaccharide composition in preparation of products for preventing and / or treating aquaculture diseases caused by vibrio parahaemolyticus, vibrio harveyi and streptococcus agalactiae, and the oligosaccharide composition is obtained by degrading polygalacturonic acid or pectin by polygalacturonase shown as SEQ ID No: 1. The degradation conditions are as follows: the concentration of the polygalacturonase is 0.045 mg / mL to 0.055 mg / mL, the concentration of the polygalacturonic acid is 7% to 9%, and the degradation time is 12 hours to 20 hours. The invention also relates to the use of the oligosaccharide composition in aquaculture. The oligosaccharide composition disclosed by the invention has good effects of inhibiting bacteria and preventing and treating aquaculture diseases.

The present invention relates to the development of a microencapsulated vaccine containing inactivated Streptococcus agalactiae, together with the adjuvants beta glucan and mineral oil, to be administered orally to fish together with feed in pelletised form or coated with the vaccine. In addition, the invention relates to the use of this vaccine to immunise fish, more specifically Nile tilapia (Oreochromis niloticus) to activate both innate and adaptive (humoral and cellular) intestinal mucosal immunity and protect tilapia against streptococcosis. Such a vaccine can be applied in intensive fishing production systems, with the aim of reducing fish mortality, reducing the use of antimicrobials and not causing environmental damage, being easy to administer and control, reducing fish handling, and being suitable for immunising large numbers of animals at the same time without stress and at low cost.

The present application relates to the field of biotechnology, and particularly relates to SNP marker combination for identifying resistance to streptococcus agalactiae disease of tilapia and application thereof. The present application discloses 11 SNP sites in UTRN gene of tilapia which are significantly related to resistance to streptococcus agalactiae disease, and constructs 18 SNP marker combinations for identifying resistance phenotype of streptococcus agalactiae disease, which can effectively improve the accuracy and genetic stability of resistance phenotype identification. In addition, the SNP typing primer and method based on MALDI-TOF mass spectrometry platform provided by the present application can realize high-throughput and high-precision genotyping, and has important application value in resistance phenotype identification and assisted selection breeding.

A method for rapidly identifying highly virulent strains of Streptococcus agalactiae, comprising: using a laser confocal Raman spectrometer to collect single-cell Raman spectra of clinical strains, and establishing a database; classifying data of III / ST17-type highly virulent GBS strains into a first group, named as GBS-HIGH, data of non-III / ST17-type GBS strains into a second group, named as GBS-OTHERS, and data of Escherichia coli into a third group, named as ECO; randomly dividing the three groups of data into a training set, a validation set and a test set in a ratio of 7:1:2; and on the basis of a ResNet18 residual neural network architecture, changing 2D convolution in a network to one-dimensional (1D) convolution suitable for a 1D Raman spectrum, determining optimal parameters by means of simulation experiments, training the weight of a model in the training set, and then selecting optimal hyperparameters by means of a result of the model on the verification set, so as to determine a trained model.

The application discloses a bocachango Mob1b antibacterial peptide, a coding sequence and use and a preparation method thereof, and relates to the following steps: based on multi-omics method, an antibacterial peptide precursor gene sequence is screened and recognized from a bocachango multi-omics data set based on biological big data, and then the bocachango Mob1b antibacterial peptide amino acid sequence is obtained through translation design, as shown in SEQ ID NO: 3; through activity simulation prediction and in-vitro experiment verification, the bocachango Mob1b antibacterial peptide shows inhibitory effects on the growth activities of streptococcus agalactiae, salmonella enteritidis, staphylococcus aureus, vibrio anguillarum and escherichia coli; in particular, the bocachango Mob1b antibacterial peptide shows inhibitory effects on the gram-positive bacteria streptococcus agalactiae, the gram-negative bacteria escherichia coli, staphylococcus aureus and vibrio anguillarum, and the inhibitory effect can be shown at a concentration of 2.5 micromoles per liter, and the best inhibitory effect can be achieved at a concentration of 5 micromoles per liter, so that the bocachango Mob1b antibacterial peptide can be used for preparing various bactericidal preparations and the like, and has good practical value and social ecological benefits.

The application discloses a kind of nanobody targeting streptococcus agalactiae SIP protein and preparation and application thereof, it is related to biotechnology field, and the nanobody that the present application is screened by nanobody phage library construction, phage screening technology, and is efficiently and specifically combined with SIP protein Nanobody, the nanobody of streptococcus agalactiae SIP protein includes VHH fragment, with the amino acid sequence shown in SEQ ID NO:1.The nanobody can be combined with streptococcus agalactiae SIP protein high specificity, and the antibody can be targeted to SIP protein by the antibody, and the application discloses a kind of nanobody targeting streptococcus agalactiae SIP protein can be applied to fish disease diagnosis and treatment.

The application provides a tilapia antibacterial peptide OnLEAP-2 and an application thereof, and an antibacterial peptide named OnLEAP-2 is cloned from tilapia, and a large amount of the antibacterial peptide is obtained through prokaryotic expression. The antibacterial peptide has a good effect of resisting Streptococcus agalactiae, so that the survival rate of the tilapia is significantly improved, and pathogens are efficiently removed. The antibacterial peptide is suitable for the prevention and treatment of bacterial diseases of economic fishes such as tilapia.

The invention discloses a method for screening crRNA based on molecular docking and application of double crRNA in improving group B streptococcus detection. A specific virulence gene in a streptococcus agalactiae genome is taken as a target spot, two potential crRNA candidates with the best detection effect are designed and screened out, and a double crRNA enhanced CRISPR detection system is established and is combined with RPA (recombinase polymerase amplification) amplification to be used for rapid and high-sensitivity detection of streptococcus agalactiae. The detection kit can complete detection from a sample to a result within one hour, the quantitative detection linear range is 103-107 cfu / mL, and the detection limit can reach 101 cfu / mL.

The present application belongs to the technical field of genetic engineering, and particularly relates to a Streptococcus agalactiae Mu-20 and a construction method and application thereof. The present application constructs a fish-derived Streptococcus agalactiae strain with a capsule polysaccharide deficiency, no hyaluronidase activity, no hemolysis and no trehalose fermentation, and the strain number is Mu-20. The strain was preserved in the Guangdong Microbial Culture Collection Center on June 13, 2022, and the preservation number is GDMCC No: 62540. The strain does not carry a chloramphenicol resistance marker gene, has high biological safety, has no risk of virulence returning to strength, has high immunogenicity, can protect immunized fish against the invasion of Streptococcus agalactiae, and is easy to cultivate, simple to use and low in production cost. The strain involved in the present application can be used to prepare a live attenuated vaccine for fish streptococcosis, and has extremely high commercial application value.

The invention discloses a freeze-dried reagent ball special for streptococcus agalactiae CRISPR detection aiming at a tilapia mossambica tissue matrix and application of the freeze-dried reagent ball, and belongs to the field of molecular diagnosis technology and on-site rapid detection of aquatic diseases. The freeze-drying reagent ball is a three-layer spherical freeze-drying body comprising an inner core layer, an isolating layer and a shell layer, the inner core layer contains an RAA amplification component aiming at a streptococcus agalactiae cfb gene, the shell layer contains a CRISPR-Cas12a detection component and a special anti-inhibition buffer system, and the isolating layer is a solid freeze-drying material layer of which the dissolution rate is slower than that of the shell layer. Through the dissolution time sequence control of the three-layer structure, the automatic sequence that an outer shell layer firstly dissolves and neutralizes an inhibitor and presets a detection system and an inner core layer then dissolves and starts amplification is realized, and multi-step reaction is integrated into one-step closed tube operation. During detection, only tilapia tissue crude extract needs to be added, and a result is interpreted through fluorescence after constant-temperature incubation. The method effectively overcomes the problems of aquatic product tissue matrix inhibition, unstable operation of an existing single-tube method and on-site pollution risk, and is high in sensitivity, extremely simple to operate and stable at normal temperature.

The invention belongs to the technical field of animal models, and particularly relates to a preparation method for constructing a rat embryo implantation failure model based on streptococcus agalactiae. The method comprises the following steps: culturing a streptococcus agalactiae strain with the number of BNCC336970 by using a solid culture medium containing sheep blood, adjusting the concentration of a bacterial solution to 1 * 10 < 8 > CFU / mL, and carrying out vaginal perfusion on an SPF-grade female SD rat every day for 21 days continuously. And after the perfusion is finished, the female mouse is mated with the male mouse at the early oestrus stage, the detected vaginal suppository is marked as the 0.5 th day of pregnancy, the mouse is killed at the 7.5 th day of pregnancy, and the model is evaluated by counting the embryo implantation quantity and detecting the expression level of endometrial tissue receptivity factors MMP9 and VEGF (Vascular Endothelial Growth Factor). A clinical pathogen pathogenic mechanism is simulated through a genital tract infection way for the first time, and the problem of physiological status deviation caused by traditional hormone intervention, immune agonists or genetic engineering methods is solved. The constructed model can stably induce the obvious reduction of the embryo implantation number and the reduction of endometrial receptivity, and provides a standardized tool for the pathological mechanism research of infectious embryo implantation failure and the screening of therapeutic drugs.

The invention provides streptococcus equi and application thereof in streptococcus equi gene editing, aims to improve the endogenous gene editing efficiency of a system on streptococcus equi, and relates to the technical field of bioengineering. According to the method, by introducing a streptococcus agalactiae RecT recombinase system, the editing efficiency of a streptococcus equi CRISPR-Cas9 gene editing system for high-yield HA is improved, and the problem that a streptococcus equi genome editing tool is low in efficiency is solved.