16 results about "Cryptosporidium parvum" patented technology

The invention belongs to the technical field of biology, and particularly relates to a cryptosporidium parvum infection immunosuppression mouse model and a construction method thereof. Aiming at the problem that the development of cryptosporidiosis drugs and vaccines is seriously limited due to the fact that no model for successfully infecting mice with cryptosporidium parvum exists at present, the invention provides the construction method of the cryptosporidium parvum infection immunosuppression mouse model, which comprises the following steps: firstly, carrying out gavage treatment on the mice for 4-7 days by adopting cyclophosphamide; constructing to obtain an immunosuppressive mouse; feeding PBS (Phosphate Buffer Solution) containing cryptosporidium parvum oocysts through the mouth, and infecting for 7-21 days; and when oocysts are continuously detected from excrement of the mouse and the content of the oocysts is higher than the infection amount, constructing the cryptosporidium parvum infection immunosuppression mouse model. The cryptosporidium parvum infection immunosuppression mouse model is successfully constructed, can be used for subsequent drug and vaccine development tests for cryptosporidium parvum diseases, and has a good application prospect.

The invention belongs to the field of biology, and discloses a cdc42 gene knockout HCT-8 cell line and application thereof, the taxonomic name of the cdc42 gene knockout HCT-8 cell line is Homo sapiens, the cdc42 gene knockout HCT-8 cell line is preserved in Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC NO: 66616; the preservation date is July 01, 2025; the preservation address is the fifth floor of building 59, No.100 courtyard, Xianlie Middle Road, Guangzhou City, Guangdong Province. According to the invention, a cdc42 gene in an HCT-8 cell is knocked out by adopting a CRISPR-Cas9 technology so as to construct a cdc42 gene knocked-out HCT-8 cell line, and the cdc42 gene knocked-out HCT-8 cell line is used as a cryptosporidium parvum cell infection model.

The invention discloses a cryptosporidium parvum AOX gene deleted strain as well as a construction method and application thereof, and the cryptosporidium parvum AOX gene deleted strain disclosed by the invention is obtained by editing an AOX gene of cryptosporidium parvum through a gene editing technology. According to the gene editing technology, the cryptosporidium parvum AOX gene is knocked out by using a CRISPR / Cas9 gene editing technology, so that a cryptosporidium parvum AOX gene deleted strain is obtained; the AOX gene is found to be related to insect toxicity for the first time, and the infection intensity of an AOX gene deleted insect strain in a gamma interferon gene knockout mouse body is remarkably reduced. Research shows that after the AOX gene is deleted from the wild type cryptosporidium parvum IIdA20G1 subtype strain, the strain has the advantage that the toxicity is obviously weakened, and can be used for preparing cryptosporidium parvum attenuated vaccines so as to prevent cryptosporidium parvum infection.

The invention discloses application of a propolis extract in preparation of a medicine for resisting Cryptosporidium parvum infection, and belongs to the technical field of biomedicine.The preparation method of the propolis extract comprises the steps that a propolis raw material is added into an extraction solvent according to the mass-to-volume ratio (g / mL) of 1: (5-20), extraction is conducted for 24-72 h at the temperature of 20-60 DEG C, then filtration is conducted, and the propolis extract is obtained. And concentrating the filtrate under reduced pressure, and drying to obtain the propolis extract. In a cryptosporidium parvum infected mouse model, the propolis extract shows a remarkable treatment effect; the propolis extract disclosed by the invention can play a role in resisting cryptosporidium parvum through a multi-target and multi-channel mechanism, has multiple biological effects of resisting parasites, inflammation, oxidation, immunoregulation, intestinal barrier protection and the like, and has good development potential and clinical application prospect.

The present invention relates to a combination vaccine comprising the antigens: a) Cryptosporidium parvum gp40; b) inactivated bovine rotavirus; c) inactivated bovine coronavirus; and d) E. coli fimbrial adhesins F5 and F41. In addition, the invention relates to methods for the preparation of the vaccine, methods for vaccinating ruminants against infection by Cryptosporidium parvum, bovine rotavirus, bovine coronavirus and E. coli, and to a kit-of-parts at least comprising a container comprising the at least one antigen according to the invention.

The application discloses application of a micro cryptosporidium cgd6_660 protein as an oocyst outer wall marker protein, and discloses that the inventors find that the cryptosporidium cgd6_660 protein is an oocyst outer wall protein, and the protein is located on the outer surface of the cryptosporidium oocyst wall. The inventors confirm the feasibility of the protein as a detection antigen. The antibody of the protein recombinant protein (as shown in a sequence table SEQ ID N0.2) can be used for immunological detection of unbroken cryptosporidium oocysts (live cryptosporidium oocysts), and the application further provides a specific polypeptide of the protein.

Cryptosporidium parvum is a highly prevalent zoonotic and anthroponotic protozoan parasite that causes a diarrheal syndrome in children and neonatal livestock, culminating in growth retardation and mortalities. Disclosed herein are inhibitors against the enzymatic activity of recombinant CpLDH protein that were identified. The inhibitors were tested for anti-Cryptosporidium effect using in vitro infection assays of HCT-8 cells monolayers. Compounds NSC158011 and NSC10447 were identified to inhibit the proliferation of intracellular C. parvum in vitro, with IC50 values of 14.88 and 72.65 μM, respectively. At doses tolerable in mice, both NSC158011 and NSC10447 significantly reduced the shedding of C. parvum oocysts in infected immunocompromised mice's feces and prevented intestinal villous atrophy as well as mucosal erosion due to C. parvum. These findings have unveiled anti-Cryptosporidium drug candidates that can be explored further for the development of therapeutic agents against C. parvum infections.

Compositions for preventing or decreasing cryptosporidiosis in animals are disclosed herein. The compositions include sources of hydrogen peroxide and are fed to animals. The animals may be neonatal calves and the cryptosporidiosis may be caused by Cryptosporidium parvum. Methods of preventing or decreasing cryptosporidiosis in animals by feeding the hydrogen peroxide-generating compositions to the animals are also disclosed. Some disclosed methods reduce the number of Cryptosporidium oocysts shed by an infected animal or reduce the infectivity of Cryptosporidium oocysts shed by an infected animal. Some methods include feeding the hydrogen peroxide-generating compositions to animals ultimately afflicted with cryptosporidiosis, and the animals gain weight despite the infection.

The application provides a double-PCR differential detection method for Cryptosporidium parvum and Eimeria bovis, which is used for synchronously differentiating and detecting Cryptosporidium parvum and Eimeria bovis in a to-be-detected sample; the double-PCR differential detection method uses the following primer combination: an upstream primer Cpar-F for Cryptosporidium parvum, a downstream primer Cpar-R for Cryptosporidium parvum, an upstream primer Ebov-F for Eimeria bovis, and a downstream primer Ebov-R for Eimeria bovis. The application also provides application of the double-PCR detection method and a kit for synchronously differentiating Cryptosporidium parvum and Eimeria bovis. The double-PCR differential detection method can simultaneously and specifically amplify two target fragments in the same reaction tube, so that rapid differentiation of two pathogens and mixed infection of the two pathogens can be realized.

The invention discloses a cryptosporidium parvum sporozoites activity detection method and device and application, and belongs to the technical field of biological detection and drug screening. The method comprises the following steps: providing a to-be-detected sample containing dead or live cryptosporidium parvum sporozoites; diluting the to-be-detected sample in a multiple ratio gradient manner and / or performing freeze thawing treatment, and adding the to-be-detected sample into a detection container; an ATP bioluminescence detection reagent is added into the detection container for oscillation treatment, so that cryptosporidium parvum sporozoite is split to release ATP, and a reaction is carried out to generate a luminescence signal; collecting the light-emitting signal, and quantitatively evaluating the survival rate or activity of the cryptosporidium parvum sporozoite according to the intensity of the light-emitting signal. The method provided by the invention is based on the ATP bioluminescence principle, can directly, quickly and quantitatively evaluate the activity of the sporozoite stage in a short time, and can be used for high-throughput screening of large-scale compound drug libraries.

The invention discloses a lactobacillus johnsonii YS my4 strain with a cryptosporidium infection resisting effect and application of the lactobacillus johnsonii YS my4 strain. The preservation number of the YS my4 strain is CCTCC (China Center For Type Culture Collection) M 20253011. The strain disclosed by the invention has the characteristics of cholate resistance, intestinal juice resistance, strong adhesiveness and capability of inhibiting salmonella and staphylococcus aureus. In addition, the strain has no hemolytic rings, and the strain has no influence on body indexes and can increase the body weight after gavage of suckling rats, so that the strain is proved to be high in safety and can be applied to the fields of medicine and food. According to the invention, the YS my4 strain is intragastrically fed to a pseudo-sterile suckling mouse infected by cryptosporidium parvum, and it is found that the strain can activate immune defense of the intestinal tract of a host by improving the MAPK phosphorylation level of the host in the state of cryptosporidium parvum infection, so that the insect loading amount of the cryptosporidium parvum is remarkably reduced; meanwhile, the YS my4 strain can improve the expression level of tight junction proteins Occludin and CLDN4, relieve pathological damage of ileum tissues and improve the intestinal barrier function, and has important application value in prevention and treatment of cryptosporidium parvum infection.

The invention belongs to the biological field and discloses a cdc42 Gene knockout HCT-8 cell line and its application, cdc42 The taxonomic name of the knockout HCT-8 cell line is Homo sapiens , deposited in Guangdong Provincial Microbial Culture Collection Center, with the deposit number GDMCC NO: 66616; deposit date July 1, 2025; deposit address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Province. The present invention uses CRISPR-Cas9 technology to knock out HCT-8 cells cdc42 Gene to construct cdc42 The gene-knockout HCT-8 cell line was used as a model for Cryptosporidium parvum cell infection.

The application discloses a Cryptosporidium parvum CpDHR1 gene replacement strain and a construction method and application thereof. The application researches and finds that the CpDHR1 gene of Cryptosporidium parvum is related to virulence, the CpDHR1 gene of a high-virulence strain is replaced by the CpDHR1 gene of a low-virulence strain through a gene editing technology, and a Cryptosporidium parvum CpDHR1 gene replacement strain is successfully constructed. Research shows that after the CpDHR1 gene of a high-virulence wild-type Cryptosporidium parvum IIdA20G1 subtype strain is replaced, the strain can still be cultured in vitro, but the in-vitro growth of the strain is significantly reduced, the virulence of the strain is weakened, the oocyst excretion intensity of the strain is weakened, the oocyst load in mice is reduced, the clinical symptoms of the mice are alleviated, the survival time of the mice is prolonged, the virulence of the strain is significantly weakened, the strain can be used for preparing a Cryptosporidium parvum low-virulence vaccine product, and Cryptosporidium parvum infection can be better prevented.

Cryptosporidium parvum is a highly prevalent zoonotic and anthroponotic protozoan parasite that causes a diarrheal syndrome in children and neonatal livestock, culminating in growth retardation and mortalities. Disclosed herein are inhibitors against the enzymatic activity of recombinant CpLDH protein that were identified. The inhibitors were tested for anti-Cryptosporidium effect using in vitro infection assays of HCT-8 cells monolayers. Compounds NSC158011 and NSC10447 were identified to inhibit the proliferation of intracellular C. parvum in vitro, with IC50 values of 14.88 and 72.65 μM, respectively. At doses tolerable in mice, both NSC158011 and NSC10447 significantly reduced the shedding of C. parvum oocysts in infected immunocompromised mice's feces and prevented intestinal villous atrophy as well as mucosal erosion due to C. parvum. These findings have unveiled anti-Cryptosporidium drug candidates that can be explored further for the development of therapeutic agents against C. parvum infections.

The invention provides application of a compound in preparation of a medicine for resisting cryptosporidium parvum, and belongs to the technical field of application of chemical raw material medicines. The compound is 5-amino-3-(1-cyano-2-(3-phenyl-4-vinylpyrazole))-1-phenyl-4-cyanopyrazole, the structural formula of the compound is shown as a formula (I): # imgabs0, and it is proved that the compound serves as an inhibitor of cryptosporidium parvum alternating oxidase CpAOX, has a remarkable effect in the aspect of preventing and treating cryptosporidium infection and is worthy of clinical popularization.

The invention discloses a multiple PCR (Polymerase Chain Reaction) primer group and a detection method for simultaneously detecting four kinds of animal-and-human protozoa of goats, which are used for detecting enterosporidium pichia, cystic protozoa, cryptosporidium parvum and giardia duodenalis. The primer group comprises the following primers: a specific primer of enterosporidium pichia, a specific primer of cyst protozoa, a specific primer of cryptosporidium parvum and a specific primer of giardia duodenalis. In addition, the invention also discloses a multiple PCR (Polymerase Chain Reaction) detection method for the enterosporidium pichia, the cystozoon, the cryptosporidium parvum and the giardia duodenalis. Experiments prove that the method has the advantages of rapidness, sensitivity and high specificity, the operation steps of traditional step-by-step PCR are remarkably reduced, the purpose of simultaneously detecting a plurality of pathogen target genes is achieved through one-time reaction, the detection cost is saved while the detection time is shortened, a foundation is laid for accurately and rapidly identifying pathogens, and the method has a relatively great development prospect.