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Cryptosporidium  spp. and  Enterocytozoon bieneusi  are two prevalent opportunistic pathogens in humans and animals. Currently, few data are available on genetic characterization of both pathogens in rabbits in China. The aim of the present study was to understand prevalence and genetic characterization of  Cryptosporidium  spp. and  E. bieneusi  in rabbits. We collected 215 fecal samples from 150 Rex rabbits and 65 New Zealand White rabbits on two different farms in Heilongjiang Province, China.  Cryptosporidium  spp. and  E. bieneusi  were tested by polymerase chain reaction (PCR) and sequencing the partial small subunit of ribosomal DNA (SSU rDNA) and the internal transcribed spacer (ITS) region of rDNA, respectively.  Cryptosporidium  was detected in 3.3% (5/150) of Rex rabbits and 29.2% (19/65) of New Zealand White rabbits. All the 24  Cryptosporidium  isolates were identified as  C. cuniculus .  Enterocytozoon bieneusi  was only found in 14.7% (22/150) of Rex rabbits. Five known genotypes: CHN-RD1 ( n  = 12), D ( n  = 3), Type IV ( n  = 2), Peru6 ( n  = 1), and I ( n  = 1), and three novel ones CHN-RR1 to CHN-RR3 (one each) were detected. By analyzing the 60-kDa glycoprotein ( gp 60) gene sequences of  C. cuniculus  isolates, three subtypes were obtained: VbA28 ( n  = 2), VbA29 ( n  = 16), and VbA32 ( n  = 3). All these three  C. cuniculus  subtypes were reported previously in humans. Four known  E. bieneusi  genotypes have been found to be present in humans. The three novel ones fell into zoonotic group 1. The results suggest zoonotic potential of  C. cuniculus  and  E. bieneusi  isolates in rabbits.

Subtyping ofCryptosporidium cuniculusand genotyping ofEnterocytozoon bieneusiin rabbits in two farms in Heilongjiang Province, China