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

Abstract

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