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Genotyping of Toxoplasma Gondii from Pig in Some Areas of Henan Province and the Culture of T.gondii RH Strain in Vitro

Author: RenQingE
Tutor: ZhangLongXian；NingChangShen
School: Henan Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Pig Toxoplasma Cell culture Multiplex PCR Microsatellite loci analysis Genotyping
CLC: S852.72
Type: Master's thesis
Year: 2009
Downloads: 75
Quote: 0
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Abstract


Toxoplasmosis is caused by Toxoplasma gondii (Toxoplasma gondii) beast of a man suffering from a protozoan. Toxoplasma gondii a wide range of host species, the common animals and humans can be infected with the disease, the most susceptible pigs in the livestock and the high morbidity and mortality. Swine Toxoplasmosis more due to the site of infection, clinical manifestations, diagnosis difficult, bring more difficulties to early diagnosis and effective prevention. T. gondii has a wide host range in different Toxoplasma animals may have different biological characteristics, so the study the of Toxoplasma gondii molecular genetics and molecular epidemiology of the diagnosis and prevention of toxoplasmosis important. Early classification of Toxoplasma gondii isolates only Toxoplasma strength of virulence for mice divided into virulent and attenuated strain. With the development of the parasite molecular genetics, has been developed to study their genotype phenotype from the study of Toxoplasma gondii. To study Toxoplasma classification from gene level distribution and virulence characteristics has initially formed will be divided into more than one genotype of Toxoplasma gondii strains perspective, and gradually worked out in a series of studies Toxoplasma genotype in different regions as well as the distribution law of the host. Diagnosis of Toxoplasma gondii from the molecular level will be to provide information for further research, and the Prevention of toxoplasmosis Toxoplasma molecular genetics. Order to grasp the the Henan Province swine toxoplasmosis infection of various outpatient and pig farms in Henan Province, Zhengzhou, Xinyang, Zhumadian, Jiaozuo and Luoyang, Nanyang region collected 1647 were feeding of suspected Toxoplasma pigs disease, application My laboratory established nested PCR molecular epidemiological investigation, the method is mainly used for the detection of Toxoplasma gondii B1 gene. To find positive cases diagnosed by PCR method. Conclusion: Looking to the 31 positive strains of insects, the positive rate was 1.88%, and are from piglets. 2. Invasion of the cells and their immune evasion mechanism carried out under laboratory conditions in order to facilitate in-depth research, we have carried out research MDCK and BHK-21 cells cultured Toxoplasma. Were removed every 12h after infection coverslips in petri dishes Giemsa staining, xylene, Toxoplasma gondii in vitro proliferation observed under the microscope. Toxoplasma gondii host cell selectivity is not strong, can be infected MDCK and BHK-21 cells, and seven days after infection, the breeding season. Genotypes of of Henan Province swine Toxoplasma gondii isolates from the molecular level to understand the author chooses five microsatellite loci: TuB2, W35, TgM-A, B18 and B17, the test isolates and international standard strain RH strain multiplex PCR amplification and sequencing of the PCR product, understand microsatellite repeat sequences after standard genotype analysis, and finally determine the genotype of Henan Province the pig source Toxoplasma isolates. Microsatellite loci analysis results show that the RH strain for genotype I, consistent with the foreign reports. NY-5, NY-8, NY-9, NY-11, NY-13 and NY-16 strains of TUB2 gene repeat sequence (TG) 9 TG repeat number different from the previously reported. W35 a repetitive sequence of the gene locus is (TC) 4CC (TC) 2 (TG) 2, and is similar to the case of atypical type. Comprehensive analysis of five microsatellite loci, six gondii strains belong to a new genotype. Study proved pig source Toxoplasma in Henan Province contains new genotypes.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Livestock parasitology > Veterinary protozoan school
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