Predomination and new genotypes of Enterocytozoon bieneusi in captive nonhuman primates in zoos in China: high genetic diversity and zoonotic significance.

Md Robiul Karim,Tongyi Li,Changshen Ning,Haiju Dong,Dezhong Li,Xiaofeng Li,Rongjun Wang,Longxian Zhang,Farzana Islam Rume,Fuchang Yu,Junqiang Li,Shouyi Li,Joao Inacio

doi:10.1371/journal.pone.0117991

Abstract

To appreciate the genetic diversity and zoonotic implications of Enterocytozoon bieneusi in nonhuman primates (NHPs) in zoos, we genotyped E. bieneusi in captive NHPs in seven zoos located at six major cities in China, using ribosomal internal transcribed spacer (ITS)-based PCR and sequence analyses. A total of 496 fecal specimens from 36 NHP species under nine families were analyzed and E. bieneusi was detected in 148 (29.8%) specimens of 25 NHP species from six families, including Cercopithecidae (28.7%), Cebidae (38.0%), Aotidae (75.0%), Lemuridae (26.0%), Hylobatidae (50.0%) and Hominidae (16.2%) (P = 0.0605). The infection rates were 29.0%, 15.2%, 18.2%, 37.3%, 29.2%, 37.7% and 44.8% in Shijiazhuang Zoo, Wuhan Zoo, Taiyuan Zoo, Changsha Wild Animal Zoo, Beijing Zoo, Shanghai Zoo and Shanghai Wild Animal Park, respectively (P = 0.0146). A total of 25 ITS genotypes were found: 14 known (D, O, EbpC, EbpA, Type IV, Henan-IV, BEB6, BEB4, Peru8, PigEBITS5, EbpD, CM1, CM4 and CS-1) and 11 new (CM8 to CM18). Genotype D was the most prevalent one (40/148), followed by CM4 (20/148), CM1 (15/148), O (13/148), CM16 (13/148), EbpC (11/148). Of them, genotypes D, EbpC, CM4 and O were widely distributed in NHPs (seen in 9 to 12 species) whereas genotypes CM1 and CM16 were restricted to one to three NHP species. In phylogenetic analysis, 20 genotypes (121/148, 81.8%), excluding genotypes BEB4, BEB6, CM9, CM4 and CM18, belonged to group 1 with zoonotic potential. New genotype CM9 clustered in group 2 with BEB4 and BEB6. The remaining two genotypes CM4 and CM18 formed new cluster (group 9) in between two other genotypic clusters found in primates. The findings of high diversity in E. bieneusi genotypes and their zoonotic potentiality concluded the importance of captive NHPs as reservoir hosts for human microsporidiosis.

Highlights

Enterocytozoon bieneusi, the dominant member of the human pathogenic microsporidian species, is a unicellular organism that infects the enterocytes of the small intestine and causes diarrhea and enteric diseases in humans, and domestic and wild animals [1,2,3]
Twenty five of 36 nonhuman primates (NHPs) species under the families were found to be infected with E. bieneusi with the occurrences ranging from 4.4 to 100%. (Table 2)
E. bieneusi was detected in 29.8% (148/496) of the NHP fecal specimens analyzed, which illustrates its common occurrence in NHPs

Summary

Introduction

Enterocytozoon bieneusi, the dominant member of the human pathogenic microsporidian species, is a unicellular organism that infects the enterocytes of the small intestine and causes diarrhea and enteric diseases in humans, and domestic and wild animals [1,2,3]. Since its first identification in AIDS patient by Desportes and others in 1985, molecular studies in various hosts and water bodies revealed E. bieneusi as potentially zoonotic pathogen worldwide [1,2,3,6,7,8,9,10,11,12,13,14,15,16,17]. With constant identification of new ITS genotypes, far, over 200 E. bieneusi genotypes have been reported in various animals, humans and water bodies worldwide [1,2,3,16,17,22,23,24,25]. Due to likely transmission between humans and animals, the large number of genetically related genotypes with broad host ranges form group 1 of zoonotic potentials. The remaining groups 7 and 6 containing the genotypes those are found in AIDS patients in Nigeria, and urban wastewater in China, respectively [3,9,16,21]

Methods

Results

Conclusion