Enterocytozoon bieneusi subsp. sequences

Ding, Hao, Zhao, Aiyun, Wang, Lingyun, Gao, Na, Sun, Yangang, Li, Junqiang & Qi, Meng, 2020, Genotypes and zoonotic potential of Enterocytozoon bieneusi in edible bullfrogs (Lithobates catesbeiana) in China, International Journal for Parasitology: Parasites and Wildlife 11, pp. 103-107 : 104

2.4. E. bieneusi sequences and phylogenetic analyses

Positive secondary PCR amplicons were sequenced by a commercial sequencing company (GENEWIZ, Suzhou, China). The sequence accuracy was confirmed via bidirectional sequencing, and the sequences obtained were aligned using ClustalX 2.1 (http://www.clustal.org/) with the reference sequences downloaded from GenBank (https:// www.ncbi.nlm.nih.gov/genbank/) to determine the species and genotypes. Representative genotypes of the nucleotide sequences we obtained were submitted to GenBank at the National Center for Biotechnology Information under accession numbers MN758739–MN758760.

Bayesian inference (BI) and Monte Carlo Markov chain methods were used to construct phylogenetic trees in MrBayes (version 3.2.6) (http://nbisweden.github.io/MrBayes/). The posterior probability values were calculated by running 1,000,000 generations. A 50% majority-rule consensus tree was constructed from the final 75% of the trees generated via BI. Analyses were run three times to ensure convergence and insensitivity to priors.