| Fascioliasis hepatica is a significant foodborne waterborne parasitic disease caused by the Fasciola hepatica parasitizing liver and bile ducts.According to statistics,there are about 250 million-300 million cattle and sheep infected with F.hepatica and cause about 3 billion USD economic losses per annum.It is particularly important that F.hepatica can also infect people.It not only brings great economic losses to animal husbandry production,but also brings great threat to human health.At present,the method of manure and autopsies is the main diagnosis method of bovine and sheep infected Fascioliasis hepatica in China.However,both of them can not be suitable for early diagnosis in vivo.In recent years,some immunological diagnostic methods have been developed,but most of these are not used on a large scale in clinical.Although there are commercialized ELISA diagnostic kits abroad,but its price is very expensive,production is relatively difficult.Therefore,we urgently need an early,sensitive,unique,and relatively low cost diagnostic method,to lay the foundation for its wide application.The development of omics technology is in full swing,and metabolomics as a new technology also plays an important role in the study of disease pathogenesis and diagnosis.And when the body is infected with pathogens,metabolic changes are also the most sensitive.In this study an artificial infection model of F.hepatica in sheep was established firstly.Collecting eggs of F.hepatica from gallbladder hatched for miracidium.Make Galba pervia collected from wild,the intermediate host,infected with miracidium and feed them for metacercariae.Feed 220 metacercariae to 11 test group sheep,and 10 sheep were bred as control group.Collect serum in zero day,3th,7th,20 th,34th,48 th,56th,72 th,84th,98 th days and plasma in 7th,56 th,98th days after infected.Determine the time of the F.hepatica maturity through the manure test,and collecting worms after slaughtering.Analyze and discus the differential metabolites of those three periods,metabolism changes of sheep and biomarkers with diagnostic potential.Serum samples were performed bio-chemistry checking 35 indexes to analyze the dynamic change process.The plasma samples were preformed metabolomics analyze based on liquid chromatography / mass spectrometry technology.The results showed that: 1)the miracidium escaped eggs as early as the 12 nd day of hatching in 37?.Cercariae broken out of the snail body as early as the 25 th day of maintaining the orders of 25-30? and in over 70% of humidity.Found eggs from sheep fecal at 84 th day of feeding metacercariae,F.hepatica recovery rate up to 50%?109/220?-60%?132/220?.2)In 35 bio-chemistry indexes were changed,including 15 liver – related,2 kidney – related and LDH,AMY.Maybe the damage was focused on liver and pancreas,and followed as the progress of F.hepatica migration.3)In ESI+mode,there were 13,27,82 differential metabolites in 7th,56 th and 98 th day,respectively.In ESI-mode,there were 0,37 and 83 differential metabolites in 7th,56 th and 98 th day,respectively.The results of metabolism pathway analysis showed that the differential metabolism pathways has the largest number and difference in 98 th day.The differential metabolism pathway in 56 th day were less.There was only one differential metabolism pathway of 7th day,it is purine metabolism.The AUC value of 7th,56 th,98th day were 1,0.964,1,respectively,showed good diagnosis ability.Conclusion: In this study,an artificial infection model of F.hepatica was established successfully.The revegetation of the cycle from eggs to adult was completed,and the material of F.hepatica was enriched.The damage to the body is always concentrated in the liver and pancreas,and the dynamic changes of biochemistry indexes and metabolomics are related to the migration process of F.hepatica.For the first time,the metabolism of sheep infected with F.hepatica was analyzed using LC/MS,and differences in plasma metabolites were found at the dpi of 7th,56 th and 98th. |
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