Metacercariae excyst in the duodenum of the host, migrate into the bile duct and then develop into adult worms. sinensis mainly by the consumption of raw or undercooked freshwater fish containing infective metacercariae. It is estimated that nearly 15 million people are infected and over 200 million people worldwide are under threat of C. subtilis spore expressing CsPmy on the surface was a promising, safe and needle-free vaccination strategy against clonorchiasis.Ĭlonorchiasis, caused by Clonorchis sinensis, is one of the most common zoonoses and is of major socio-economic importance in East Asia, including China, East Russia, Korea and Vietnam. Although both intragastric and intraperitoneal immunization elicited a similar protective effect, intragastric immunization induced stronger mucosal immune response without side effects to the liver, intestine and gut microbiota, compared with intraperitoneal immunization. Both intragastrical and intraperitoneal immunization with B.s-CotC-CsPmy spores induced systemic and local mucosal immune response in mice. ConclusionsĬsPmy expressed on the spore surface maintained its immunogenicity. Liver fibrosis degree in B.s-CotC-CsPmy spores treated groups was also significantly reduced. Intragastric and intraperitoneal immunization of B.s-CotC-CsPmy spores in mice resulted in egg reduction rates of 48.3 and 51.2% after challenge infection, respectively. Additionally, the diversity and abundance of gut microbiota were not changed after oral immunization. No inflammatory injury was observed in the intestinal tissues and there was no significant difference in levels of enzyme-indicated liver function among the groups. Furthermore, numerous IgA-secreting cells were detected in intestinal mucosa of intragastrically immunized mice. The sIgA level in intestinal mucus, feces and bile of B.s-CotC- CsPmy orally treated mice were also significantly raised. Specific IgG in sera and intestinal mucus were increased after intraperitoneal and intragastrical immunization. ResultsĬsPmy was successfully expressed on the spore surface and the fusion protein on the spore surface with thermostability. Additionally, biochemical enzyme activities in sera, the intestinal histopathology and gut microflora of spore-treated mice were investigated. The immune response and protective efficacy of the recombinant spore were investigated in BALB/c mice after intragastrical or intraperitoneal immunization. We constructed a recombinant plasmid of PEB03-CotC-CsPmy, transformed it into Bacillus subtilis WB600 strain ( B.s-CotC- CsPmy), and confirmed CsPmy expression on the spore surface by SDS-PAGE, Western blotting and immunofluorescence assay. sinensis (CsPmy) is a potential vaccine candidate. Previous studies indicated that paramyosin of C. Effective prevention strategies are urgently needed to control this food-borne infectious disease. Clonorchiasis caused by Clonorchis sinensis has become increasingly prevalent in recent years.

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