Abstract
Enterocytozoon bieneusi is the most common pathogen of microsporidian species infecting humans worldwide. Although E. bieneusi has been found in a variety of animal hosts, information on the presence of E. bieneusi in captive kangaroos in China is limited. The present study was aimed at determining the occurrence and genetic diversity of E. bieneusi in captive kangaroos. A total of 61 fecal specimens (38 from red kangaroos and 23 from grey kangaroos) were collected from Nanjing Hongshan Forest Zoo and Hongshan Kangaroo Breeding Research Base, Jiangsu province, China. Using the nested PCR amplification ITS gene of rRNA of E. bieneusi, totally 23.0% (14/61) of tested samples were PCR-positive with three genotypes (i.e. one known genotype, CHK1, and two novel genotypes, CSK1 and CSK2). Multi-locus sequence typing using three microsatellites (MS1, MS3, and MS7) and one minisatellite (MS4) revealed one, five, two, and one types at these four loci, respectively. In phylogenetic analysis, the two genotypes, CHK1 and CSK1, were clustered into a new group of unknown zoonotic potential, and the novel genotype CSK2 was clustered into a separate clade with PtEb and PtEbIX. To date, this is the first report on the presence of E. bieneusi in captive red kangaroos in Jiangsu province, China. Furthermore, a high degree of genetic diversity was observed in the E. bieneusi genotype and seven MLGs (MLG1-7) were found in red kangaroos. Our findings suggest that infected kangaroo may act as potential reservoirs of E. bieneusi and be source to transmit infections to other animal.
Highlights
Enterocytozoon bieneusi is an unicellular enteric microsporidian that causes diarrhea in a variety of domestic and wild animals worldwide as well as in humans, especially in children and immunosuppressed individuals [1]
To better assess the diversity of the E. bieneusi genotypes in the present study and to determine the genetic relationship between the novel ones isolated from the kangaroos and the reference sequences previously published in GenBank, phylogenetic analysis was performed by constructing a neighboring-joining tree using Mega 6 software, which is based on evolutionary distances calculated using a Kimura 2-parameter model
14 (23.0%) out of 61 captive kangaroos were found to be infected with E. bieneusi, and all the positive specimens belonged to red kangaroos from the Hongshan Kangaroo Breeding Research Base, which acts as the largest kangaroo breeding zoo in China
Summary
Enterocytozoon bieneusi is an unicellular enteric microsporidian that causes diarrhea in a variety of domestic and wild animals worldwide as well as in humans, especially in children and immunosuppressed individuals [1]. Enterocytozoon bieneusi in captive red kangaroos in China More than 240 genotypes of E. bieneusi have been identified based on the analysis of the sequences of the single internal transcribed spacer (ITS) region of the rRNA gene [5,6,7,8].
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