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Background   Cryptosporidium  spp. and  Giardia duodenalis  are major intestinal pathogens that can cause diarrheal diseases in humans, especially children.  Enterocytozoon bieneusi  is another parasite which can cause gastrointestinal tract disorders, with diarrhea being the main clinical symptom. However, few genetic studies of these parasites in pediatric inpatients in China have been published.    Methods  To assess the genetic characteristics and epidemiological status of these parasites, a total of 2284 fecal samples were collected from children in the pediatric departments of three hospitals in Zhengzhou, central China, and screened for these protozoans with PCR, based on the small subunit ribosomal RNA ( SSU  rRNA) genes of  Cryptosporidium  spp. and  G. duodenalis  and the internal transcribed spacer (ITS) of  E. bieneusi .    Results  Six (0.26%), 14 (0.61%), and 27 (1.18%) of the samples were positive for  Cryptosporidium  spp.,  G. duodenalis  and  E. bieneusi , respectively. Of the 12 successfully sequenced  G. duodenalis  isolates, four were identified as assemblage A and eight as assemblage B. In subtype and multilocus genotype (MLG) analyses,  C. parvum  IIdA19G1 ( n  = 4) and two novel  G. duodenalis  MLGs belonging to subassemblage AII ( n  = 3) and BIV ( n  = 5) were successfully identified. The  E. bieneusi  isolates included genotypes D ( n  = 17), J ( n  = 2), PigEBITS7 ( n  = 1), BEB6 ( n  = 1), and CM8 ( n  = 1). This is the first report of  C. parvum  subtype IIdA19G1 in HIV-negative children and  E. bieneusi  genotype CM8 in humans.    Conclusions  The dominance of zoonotic  C. parvum  subtype IIdA19G1 indicates that this parasite is turning into zoonotic origin from human-to-human transmission. The phylogenetic analysis also revealed the zoonotic origins and anthroponotic transmission potential of  G. duodenalis  and  E. bieneusi , suggesting more efforts must be made to minimize the threat these pathogens pose to public health.

Molecular characterization of three intestinal protozoans in hospitalized children with different disease backgrounds in Zhengzhou, central China