| ObjectiveClonorchis sinensis is a parasitewhich seriously harmful to human health,and is also a food-borne parasite that focuses on prevention and control of China.The parasite can cause liver lesions,infection and even liver cancer of the host.Hunan province is an endemic province of Clonorchis sinensis.We intend to clarify the prevalence of Clonorchis sinensis in Hunan Province,its impact on the host,genome polymorphism and evolutionary mutation through continuous monitoring and investigation,establishment of experimental animal model,genome sequencing and other methods.A systematic study of the epidemiology and genomics of Clonorchis sinensis in Hunan province has enabled us to have a deep understanding of the interaction and adaptation between Clonorchis sinensis and intermediate and final hosts as well as environmental changes at the cellular and molecular levels,thus guiding the in-depth study of the host-pathogen ecological relationship.It will also promote the update of prevention and control methods and tools for important food-borne parasitic diseases,and lay a foundation for the development of new diagnostic and therapeutic strategies,prevention and control methods and methods.Method1.Epidemiological investigation of Clonorchis sinensis was carried out in 122 counties(districts)in Hunan Province,and the infection rate of Clonorchis sinensis was investigated by Kato-Katz method.2.The early disease model of Clonorchis sinensis was established by using newly weaned SD rats.The growth,feeding,blood index,biochemical index,viscera development,intestinal flora and histopathology of animals were systematically studied.3.Cox1 gene and Nad1 gene was used as amplification sites,a molecular biological method was established to detect the metacercariae of Clonorchis sinensis in fish muscle.And the molecular biological detection method was compared with the traditional muscle compression microscopy method.4.Clonorchis sinensis collected from Tongdao County,Lengshuitan District and Qiyang County were amplified with mitochondrial genes Cox1 gene and Nad1 gene as the amplification sites.The amplified products were sequenced and the consanguinity was analyzed.5.The whole genome of clonorchis sinensis isolates from Tongdao County was sequenced,and the sequencing results were compared with the standard Clonorchis sinensis from Gen Bank.Results1.Except For Yongzhou and Huaihua,the infection rate of population in other parts of the province was below 1.5%.The infection rate of Carp dorsal fin was 99.44% in Qiyang.The Population infection rate was 26.90% in Tongdao County,18.96% in Qiyang County and21.80% in Lengshuitan.2.SD rats infected with Clonorchis sinensis,liver hypertrophy and hyperemia were observed to varying in all dose infection groups,and weight increased,forming a significant drug-dose response relationship.No significant changes were observed in other tissues.The increase of white blood cells,mainly eosinophils,showed a dose-dose reaction relationship,the difference was significant.Aspartate aminotransferase and alanine aminotransferase were increased.The number of bifidobacterium and Lactobacillus in intestinal tract decreased significantly,while the number of enterococcus,Enterobacter and Clostridium perfringens increased.3.The detection rate of cyscariae of Clonorchis sinensis by muscle compression microscopy was 75.31%,Cox1 gene was 96.30%,Nad1 gene was 94.44%,suggesting that the detection rate of molecular biological detection method was better than that of traditional microscopy method,and the difference was statistically significant.4.There was high homology between Qiyang county and the Lengshuitan District,and the genetic relationship was closed between the two areas and guangdong province.The homology of the species in Tongdao County was not closed toother Clonorchis sinensis in China,but high with that in America and Russia.5.There were a large number of mutations between Clonorchis sinensis in Tongdao County and standard strains.Sample C2,showed greater genomic diversity.Genomic information showed the types of mutations,mainly SNP mutations and In Del mutations,and most of the locations and numbers of mutations occurred in the exon region.Conclusion1.Qiyang County,Lengshuitan District has always eaten raw fish,so Clonorchis sinensis infection is at a relatively high level,Tongdao high infection rate is related to the It’s underdeveloped economic status and unhealthy living habits of local residents,but also related to the lack of publicity and education.2.SD rats infected with Clonorchis sinensis,there was edema and congestion in the liver,which may be related to the inflammatory response caused by soluble antigens secreted by adults,eggs and other inflammatory factors,thus inducing cytokines such as inflammatory chemokines.However,Clonorchis sinensis infection can affect the imbalance of oxidative stress response of liver cells in mammals and the pathological changes of hepatobiliary duct tissue.The degree of liver injury was positively correlated with the load of clonorchis sinensis.After experimental SD rats were infected with Clonorchis sinensis,a series of intestinal microenvironment changes were important reasons for intestinal adverse symptoms of the host rats.This study provides new theoretical support for the diagnosis and treatment of Clonorchiasis in clinic.3.The molecular biological detection method is simple,rapid and has a high detection rate.It is suitable for the local strains of Hunan Province,and provides a new technical support for the epidemiological investigation and detection of the metacercariae in the fish of the intermediate host of Clonorchis sinensis.4.Phylogenetic analysis suggested that the species of Clonorchis sinensis in Tongdao county probably originated from high latitude Russia,which was related to species invasion and migration.These results laid a foundation for the study of detection methods,control,drugs and vaccines suitable for clonorchis sinensis in this area.5.The establishment of the Clonorchis sinensis genome database in Tongdao County has important research guidance and practical significance in the biological characteristics,genotype characteristics,population evolution,individual differences,detection antigen screening,selection of deinsectization drugs,vaccine development and other aspects of Clonorchis sinensis.This thesis contained28 figures,40 tables and 146 references. |
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