Abstract
BackgroundMost studies on Enterocytozoon bieneusi are conducted based on the internal transcribed spacer (ITS) region of the rRNA gene, whereas some have examined E. bieneusi population structures. Currently, the population genetics of this pathogen in giant panda remains unknown. The objective of this study was to determine the E. bieneusi population in captive giant pandas in China.ResultsWe examined 69 E. bieneusi-positive specimens from captive giant pandas in China using five loci (ITS, MS1, MS3, MS4 and MS7) to infer E. bieneusi population genetics. For multilocus genotype (MLG) analysis of E. bieneusi-positive isolates, the MS1, MS3, MS4, and MS7 microsatellite and minisatellite loci were amplified and sequenced in 48, 45, 50 and 47 specimens, respectively, generating ten, eight, nine and five types. We successfully amplified 36 specimens and sequenced all five loci, forming 24 MLGs. Multilocus sequence analysis revealed a strong and significant linkage disequilibrium (LD), indicating a clonal population. This result was further supported by measurements of pairwise intergenic LD and a standardized index of association (ISA) from allelic profile data. The analysis in STRUCTURE suggested three subpopulations in E. bieneusi, further confirmed using right’s fixation index (FST). Subpopulations 1 and 2 exhibited an epidemic structure, whereas subpopulation 3 had a clonal structure.ConclusionsOur results describe E. bieneusi population genetics in giant pandas for the first time, improving the current understanding E. bieneusi epidemiology in the studied region. These data also benefit future studies exploring potential transmission risks from pandas to other animals, including humans.
Highlights
Most studies on Enterocytozoon bieneusi are conducted based on the internal transcribed spacer (ITS) region of the rRNA gene, whereas some have examined E. bieneusi population structures
ITS gene analysis is widely used in molecular epidemiological studies of genetically diverse E. bieneusi, identifying over 200 genotypes [10, 11]
Multilocus genotyping (MLST) has been developed, targeting three microsatellites (MS1, MS3 and MS7) and a minisatellite (MS4) to provide high-resolution E. bieneusi genotyping based on length polymorphisms and single nucleotide polymorphisms (SNP) [13]
Summary
Most studies on Enterocytozoon bieneusi are conducted based on the internal transcribed spacer (ITS) region of the rRNA gene, whereas some have examined E. bieneusi population structures. The population genetics of this pathogen in giant panda remains unknown. The objective of this study was to determine the E. bieneusi population in captive giant pandas in China. ITS gene analysis is widely used in molecular epidemiological studies of genetically diverse E. bieneusi, identifying over 200 genotypes [10, 11]. ITS analysis is adequate for genotyping and epidemiological studies of E. bieneusi in the absence of recombination. Together with ITS analysis, multilocus genotyping has been gradually applied to E. bieneusi genetic characterization. The results contribute to the establishment of effective control measures by enabling elucidation of transmission routes, characterization of the public health consequences of animal-derived E. bieneusi isolates, and identification of the sources of human E. bieneusi infections
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