Infection and molecular characterization of Enterocytozoon bieneusi and Cyclospora cayetanensis in pet dogs and cats in Shanghai
Objective To investigate the infection and molecular characteristics of Enterocytozoon bieneusi and Cyclospora cayetanensis in pet dogs and cats in Shanghai and assess the zoonotic potential.Methods Fresh fecal samples were collected from dogs and cats at a pet hospital in Shanghai from November 2021 to June 2022.Ge-nomic DNA was extracted from the fecal samples,and nested PCR was used to amplify the internal transcribed spacer(ITS)region of the ribosomal RNA(rRNA)of E.bieneusi and the small subunit ribosomal RNA(SSU rRNA)sequence of C.cayetanensis.The positive products were subjected to bidirectional sequencing and sequence alignment by BLAST in the GenBank database.The phylogenetic tree was constructed using MEGA 11.0 software based on the neighbor-joining method.Results A total of 145 fecal samples were collected(99 from dogs and 46 from cats).The positive rates of E.bieneusi and C.cayetanensis were 4.1％(6/145)and 4.8％(7/145),respectively.The positive rates of E.bieneusi and C.cayetanensis in dogs were 3.0％(3/99)and 6.1％(6/99),respectively,while those in cats were 6.5％(3/46)and 2.2％(1/46),respectively.The ITS sequences of 6 E.bieneusi were 100％identical to human genotype A(GenBank accession number:MK982500),and the SSU rRNA sequences of 7 C.cayetanensis were 100％identical to human C.cayetanensis(GenBank accession number:KJ569533).The results of the phylogenetic tree analysis showed that all the E.bieneusi isolates belonged to the same branch as the reported human E.bieneusi genotype A isolates and fell into group 1.All the C.cayetanensis isolates belonged to the same branch as the reported human C.cayetanensis isolates.Conclusion E.bieneusi and C.cayetanensis infections were detected in pet dogs and cats in Shanghai,all of which were caused by zoonotic parasite strains.
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