Abstract
Enterocytozoon bieneusi is a common opportunistic pathogen causing diarrhea and enteric disease in a variety of animal hosts. Although it has been reported in many animals, there is no published information available on the occurrence of E. bieneusi in red-bellied tree squirrels. To understand the occurrence, genetic diversity, and zoonotic potential of E. bieneusi in red-bellied tree squirrels, 144 fecal specimens from Sichuan province, China, were examined by PCR amplification and sequencing of the internal transcribed spacer (ITS) region of the ribosomal RNA (rRNA) gene of E. bieneusi. The overall infection rate of E. bieneusi 16.7% (24/144) was observed in red-bellied tree squirrels. Altogether five genotypes of E. bieneusi were identified: three known genotypes D (n = 18), EbpC (n = 3), SC02 (n = 1) and two novel genotypes CE01, CE02 (one each). Multilocus sequence typing (MLST) analysis employing three microsatellite (MS1, MS3, MS7) and one minisatellite (MS4) revealed 16, 14, 7 and 14 positive specimens were successfully sequenced, and identified eight, three, three and two genotypes at four loci, respectively. In phylogenetic analysis, the three known genotypes D, EbpC, and SC02 were clustered into group 1 with zoonotic potential, and the two novel genotypes CE01 and CE02 were clustered into group 6. The present study firstly reported the occurrence of E. bieneusi in red-bellied tree squirrels in China, and the E. bieneusi genotypes D and EbpC were found in humans previously. These results indicate that red-bellied tree squirrels may play a potential role in the transmission of E. bieneusi to humans.
Highlights
Microsporidia, obligate intracellular eukaryotic pathogens, are composed of approximately 1300 species in 160 genera [1, 2]
DNA sequencing and subsequent analysis of the internal transcribed spacer (ITS)-PCR products from the 24 E. bieneusipositive specimens revealed the existence of three known E. bieneusi genotypes (D, EbpC, SC02), and two novel genotypes, which were named as CE01 and CE02 (Table 1)
Genotype D was the most prevalent (75.0%, 18/24), and was observed in samples from all the three cities, followed by EbpC, which was detected in three specimens from Chengdu and Ya'an cities (12.5%, 3/24); and the genotypes, SC02, CE01, and CE02 were found in one specimen each collected from Ya'an and Chengdu (4.7%, 1/24) (Table 1)
Summary
Microsporidia, obligate intracellular eukaryotic pathogens, are composed of approximately 1300 species in 160 genera [1, 2]. Human-Pathogenic Enterocytozoon bieneusi in Sichuan, China have been detected in humans [3]. Enterocytozoon bieneusi is the most prevalent microsporidian species and accounts for more than 90% of the cases of human microsporidiosis [4, 5]. Clinical symptoms caused by E. bieneusi in immunocompetent individuals are self-limiting diarrhea and malabsorption. E. bieneusi cause a life-threatening diarrhea in immune-compromised patients, in AIDS patients and organ transplant recipients [3, 6]. Apart from humans, E. bieneusi has been observed in many vertebrates species, including mammals, reptiles, and birds [7,8,9,10]
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