Molecular characterization and zoonotic potential of Entamoeba spp., Enterocytozoon bieneusi and Blastocystis from captive wild animals in northwest China.

BACKGROUND: Parasites Entamoeba spp., Enterocytozoon bieneusi and Blastocystis are prevalent pathogens causing gastrointestinal illnesses in animals and humans. Consequently, researches on their occurrence, distribution and hosts are crucial for the well-being of both animals and humans. Due to the confined spaces and frequent interaction between animals and humans, animal sanctuaries have emerged as potential reservoirs for these parasites. In this study, the wildlife sanctuary near the Huang Gorge of the Qinling Mountains in northwest China is chosen as an ideal site for parasite distribution research, considering its expansive stocking area and high biodiversity. RESULTS: We collected 191 fecal specimens from 37 distinct wildlife species and extracted genomic DNA. We identified these three parasites by amplifying specific gene regions and analyzed their characteristics and evolutionary relationships. All the parasites exhibited a high overall infection rate, reaching 90.05%. Among them, seven Entamoeba species were identified, accounting for a prevalence of 54.97%, with the highest infection observed in Entamoeba bovis. In total, 11 Enterocytozoon bieneusi genotypes were discovered, representing a prevalence of 35.08%, including three genotypes of human-pathogenic Group 1 and two novel genotypes (SXWZ and SXLG). Additionally, 13 Blastocystis subtypes were detected, showing a prevalence of 74.87% and encompassing eight zoonotic subtypes. All of the above suggests significant possibilities of parasite transmission between animals and humans. CONCLUSIONS: This study investigated the occurrence and prevalence of three intestinal parasites, enhancing our understanding of their genetic diversity and host ranges in northwest China. Furthermore, the distribution of these parasites implies significant potential of zoonotic transmission, underscoring the imperative for ongoing surveillance and implementation of control measures. These efforts are essential to mitigate the risk of zoonotic disease outbreaks originating from wildlife sanctuary.

[Biological activity of purificated swine XCL1 protein and preparation of its polyclonal antibody].

AIM: To obtain purificated his-XCL1 recombinant protein of porcine in vitro and prepare the associated polyclonal antibody; To study how the recombinant protein affects on lymphocytes proliferation. METHODS: Purify the recombinant protein using HiTrap(TM); Chelating HP chromatographic column; Check the purified product using SDS-PAGE; Detect the expression of the recombinant protein using Western blot; Immunize the experimental animals with the purified fusion protein to prepare the serum containing the associated polyclonal antibody. The serum will then undergo double immnodiffusion test and indirect ELISA test to determine the polyclonal antibody titer. Then, test the condition of lymphocytes proliferation by MTT. RESULTS: Single target strip could be seen under conducting the SDS-PAGE electrophoresis when the concentration of the binding buffer is 40 mmol/L and the concentration of the elution buffer is 500 mmol/L; Western blot test showed that the recombinant protein could be successfully expressed; Double immunodiffusion test showed the antigen-antibody binding ratio to be 1:8, and the titre of antibodies was 1:12 800 detected by indirect ELISA. The result of MTT showed that both the native XCL1 and the recombinant protein could stimulate lymphocytes proliferation, and this stimulating effect could be effectively blocked by the polyclonal antibody we prepared. CONCLUSION: To conclude, this recombinant protein has biological activity and this research can provide basic material for further investigation of the function of XCL1 in swine.