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Enterocytozoon  ( E. )  bieneusi  and  Cryptosporidium  spp. are the most important zoonotic enteric pathogens associated with diarrheal diseases in animals and humans. However, it is still not known whether  E. bieneusi  and  Cryptosporidium  spp. are carried by wild rodents in Shanxi, Guangxi, Zhejiang, Shandong, and Inner Mongolia, China. In the present study, a total of 536 feces samples were collected from  Rattus (R.) norvegicus ,  Mus musculus ,  Spermophilus  ( S. )  dauricus , and  Lasiopodomys brandti  in six provinces of China, and were detected by PCR amplification of the SSU rRNA gene of  Cryptosporidium  spp. and ITS gene of  E. bieneusi  from June 2017 to November 2020. Among 536 wild rodents, 62 (11.6%) and 18 (3.4%) samples were detected as  E. bieneusi - and  Cryptosporidium  spp.-positive, respectively. Differential prevalence rates of  E. bieneusi  and  Cryptosporidium  spp. were found in different regions.  E. bieneusi  was more prevalent in  R. norvegicus , whereas  Cryptosporidium  spp. was more frequently identified in  S. dauricus . Sequence analysis indicated that three known  Cryptosporidium  species/genotypes ( Cryptosporidium viatorum ,  Cryptosporidium felis , and  Cryptosporidium  sp. rat genotype II/III) and two uncertain  Cryptosporidium  species ( Cryptosporidium  sp. novel1 and  Cryptosporidium  sp. novel2) were present in the investigated wild rodents. Meanwhile, 5 known  E. bieneusi  genotypes (XJP-II, EbpC, EbpA, D, and NCF7) and 11 novel  E. bieneusi  genotypes (ZJR1 to ZJR7, GXM1, HLJC1, HLJC2, and SDR1) were also observed. This is the first report for existence of  E. bieneusi  and  Cryptosporidium  spp. in wild rodents in Shanxi, Guangxi, Zhejiang, and Shandong, China. The present study also demonstrated the existence of  E. bieneusi  and  Cryptosporidium  spp. in  S. dauricus  worldwide for the first time. This study not only provided the basic data for the distribution of  E. bieneusi  and  Cryptosporidium  genotypes/species, but also expanded the host range of the two parasites. Moreover, the zoonotic  E. bieneusi  and  Cryptosporidium  species/genotypes were identified in the present study, suggesting wild rodents are a potential source of human infections.

Molecular Detection of Cryptosporidium spp. and Enterocytozoon bieneusi Infection in Wild Rodents From Six Provinces in China