*2.3. MLG and Phylogenetic Analysis*

Collectively, sequence data sets from the three loci were available from 10 isolates. Multi-locus genotyping yielded 10 novel MLGs (one MLG for assemblage A and nine MLGs for assemblage B) (Table 3). The single MLG in assemblage A was identified as AI–novel 1, whereas the nine MLGs in assemblage B were designated from BIV–novel 1 to BIV–novel 7. Phylogenetic relationships of MLGs in assemblages A and B with the reference genotypes are illustrated in Figure 1. Based on the phylogenetic tree, AI–novel 1 identified in the present study clustered more closely to MLG AI-2 (Figure 1A). Within the assemblage B phylogenetic analysis, all MLGs in assemblage B were identified as BIV and clustered in the MLG BIV branch (Figure 1B).

**Figure 1.** Nucleotide neighbor-joining trees based on concatenated datasets for *bg*, *tpi*, and *gdh* gene sequences of *G. duodenalis* assemblage (**A**,**B**) isolates obtained in this study and sequences retrieved from the GenBank database. Bootstrap values greater than 50% from 1000 replicates are shown on nodes. The bold texts represent the isolates of this study.
